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Construction of Vector Containing Artificial miRNA to Specifically Inhibit MPG1 Gene Expression in Magnaporthe grisea Causing Rice Blast

Nguyen Thi Phuong Thao, Nguyen Trang Hieu, Phan Thi Huong, Nguyen Thi Thuy Linh

Date Received: 17-09-2013 / Date Accepted: 26-09-2013 / Date Published: 02-07-2025
In hot and humid climate countries, such as Vietnam, Magnaporthe grisea causes serious damage to rice. MPG1, a pathogenicity gene of Magnaporthe grisea is related to the process of aspersorium formation, symptom development and sporulation. Therefore, inhibition of MPG1 gene expression probably results in reduced or loss of pathogenicity of Magnaporthe grisea. A coding fragment of MPG1 gene of 4 rice blast fungus isolates was cloned and the sequencing results showed that the values of gene identity ranged from 99.3-100% among isolates in comparison with the control strain (Guy-11, ID genbank: L20685.2). Two artificial miRNAs were designed using natural Arabidopsis thaliana miRNA-319a backbone to specifically inhibit MPG1 gene expression. These artificial miRNAs were ligated into plant expression pPS1 vector to create gene construct for transferring into rice plant via Agrobacterium tumefaciens. 18 lines of transgenic rice variety J02 were identified to carry pre-amiR-P1 and these lines showed adaptation and survival under the greenhouse condition.
DOI: https://doi.org/10.31817/tckhnnvn.2013.11.6.
Structural Analysis and Data Mining of the Expression Profiles of Genes encoding TCP Transcription Factors in Pummelo (Citrus grandis)

Chu Dúc Hà, Nguyẽn Thu Huòng, Bùi Thị Thu Huong, La Viẹt Hòng, Le Thị Ngọc Quỳnh, Phạm Phuong Thu, Nguyẽn Van Lọc

Date Received: 10-03-2020 / Date Accepted: 04-05-2020
TCP transcription factor is considered as the key regulatory proteins, which are demonstrated to involve in the growth and development of plants. However, the information of TCP in pummelo (Citrus grandis) has been still lacking.The aims of this study were to identify and characterize the basic features of TCPs in pummelo. As the results,a total of 21 genes encoding TCP havebeen identified in the pummelo's genome. Based on various bioinformatics tools, the physic-chemical properties of TCP family in pummelo have beenfound to be very variable, hydrophilic, as confirmed in other plants-species. The majority of CgTCP genes consist ofonly one exon. We have also found that TCP in pummelo could classify into 3 plant-specific groups, including PCF1, CIN and CYC/TB1. Of our interest, according to the transcriptome atlas of the sweet orange (C. sinensis), we demonstrated that the homologous gene of CgTCP02 is strongly induced in leaf, flower and fruit. Taken together, our results could get insight into the TCP family in pummeloand initially evaluate the expression profiles of TCP-coding genes in the major organs, and thus, provide the candidate genes for further functional characterizations.
Comparison of Spectral Clustering Algorithms for Gene Expression Data

Hoang Thi Thanh Giang, Nguyen Thi Thuy Hanh, Nguyen Hoang Huy

Date Received: 22-07-2015 / Date Accepted: 03-09-2015 / Date Published: 13-06-2025
Spectral clustering algorithms have been the most effective algorithms to divide genes into groups accordingto the degree of their expression similarity. Such a grouping may suggestthat the respective genes are correlated and/or co-regulated, and subsequently indicatesthat the genes could possibly share a common biological role. In this paper,three spectral clustering algorithms were investigated: unnormalized spectral clustering, normalized spectral clustering according to Shi and Malik (2000), and normalized spectral clustering according to Ng, Jordan and Weiss (2002). The algorithms were benchmarked against each other.The performance of the three clustering algorithms wasstudied on time series expressiondata using Dynamic TimeWarping (DTW) distance in order to measure similarity betweengene expression profiles. Four different cluster validation measures were usedto evaluate the clustering algorithms: Connectivity and Silhouette Index for estimatingthe quality of clusters, Jaccard Index for evaluating the stability of a clustermethod and Rand Index for assessing the accuracy. The results were analyzedby Friedman’s test. The performance of normalized spectral clustering according to Ng, Jordan and Weiss (2002)wasdemonstratedto be the best under the Silhouetteand Rand validation indices.
DOI: https://doi.org/10.31817/tckhnnvn.2015.13.6.
CLONING, EXPRESSION AND PURIFICATION OF ABO3 GENE INVOLVED IN DROUGHT STRESS TOLERANCE FROM Arabidopsis thaliana in Escherichia coli

Nguyen Xuan Canh, Nguyen Thi Nhan, Nguyen Thi Phuong Anh, Tran Kim Oanh, Nguyen Thi Thuy Hanh

Date Received: 09-05-2016 / Date Accepted: 10-08-2016 / Date Published: 10-08-2016
Drought is one of the most severe environmental stress factors that affects the growth and development of plants. The effects of drought are foreseen to increase with climate change and growing water scarcity. In many parts of the world, including Vietnam, plants may frequently encounter drought stress. Plants have evolved specific mechanisms to respond to drought stresses. Studying these protective mechanisms will contribute to our knowledge of tolerance and resistance to stress. Recently, some studies indicated that the ABO3 gene coding for WRKY63 is a key gene in stress signalling pathways related to drought tolerance, of which the functions and interactions are currently unknown. Therefore, investigating its functions and interactions are considered effective strategies to control a plant’s resistance against drought. In this study, we show the results of expression and purification of the ABO3 protein coding for WRKY63 in E. coli, a solid step towards studying about the protein’s interactions and functions. The ABO3 gene was amplified from cDNA of Arabidopsis thaliana by PCR, cloned into T-Blunt vectors, and sequenced. This gene then was subcloned into the expression vector pQE-30 and expressed in E. coli strain M15. Finally, the ABO3 recombinant protein was initially purified by affinity chromatography for further studies.
DOI: https://doi.org/10.31817/tckhnnvn.2016.14.7.
Evaluation of Feature Selection Methods for Gene Expression Data Classifcation

Phan Thi Thu Hong, Nguyen Thi Thuy

Date Received: 11-08-2015 / Date Accepted: 08-03-2016 / Date Published: 12-06-2025
Selection of relevant genes that have effects in some diseases is a challenging task in gene expression studies. Most gene selection studies focused on assessing the association between individual gene and the disease. In fact, diseases are thought to involve a complex etiology including complicated interactions between many genes and the disease. Random Forest (RF) method has recently been successfully used for identifying genetic factors that have effects in some complex diseases. In spite of performing well in some data sets with moderate size, RF still suffers from working for selecting informative genes and building accurate prediction models. In this paper, we investigatedsome methods in learning advanced random forests that allow one to select a sub-set of informative genes (most relevant to disease). The method can therefore reduce the dimensionality and can perform well in prediction high-dimensional data sets. The performance of these methods has been analyzed for finding the robust one for each interest objective (the accuracy of the prediction model or the smallest possible set of relevant genes) based on experiments results on 8 available public data sets of gene expression from the repository of biomedical data sets (Kent Ridge) and bioinformatics data sets(Bioinformatics).
DOI: https://doi.org/10.31817/tckhnnvn.2016.14.3.
Genetic Transformation of Some Cherry Tomato (Lycopersicon esculentum Mill.) Cultivars with The HbsAg Gene

Nguyen Thi Phuong Nam, Tran Thi Ngoc Ha, Le Hoan Hao, Le Tan Duc, Pham Duc Tri, Phan Tuong Loc, Nguyen Huu Tam, Bui Minh Tri, Nguyen Huu Ho

Date Received: 27-07-2014 / Date Accepted: 24-09-2014 / Date Published: 06-08-2025
The present paper reported the results on the genetic transformation of two cherry tomato cultivars, TN412 (pureline) and TN TN48 (F1 hybrid) with the HBsAg for long-term objective of producing the plant edible vaccine against Hepatitis B virus. Shoots regenerated from cotyledons served as materials for transformation experiments. The 7-day old cotyledons of tomato seedlings were infected and co-cultivated for 2 days with the Agrobacterium tumefaciens strain carrying the HBsAg gene [driven by the PDS (phytoene desaturase) and T7 promoters], kanamycin resistance gene nptII and gusA reporter gene. Two days after co-cultivation, the cotyledons were washed with cefotaxime solution and cultured on the MS medium containing + 0.5 mg/l IAA + 2 mg/l BA + 500 mg/l cefotaxime and 30 - 50 mg/l kanamycin for selection of transformants. Through at least 04 rounds of selection (15 - 20 days/round), various kanamycin resistant (KR) lines were obtained in both cultivars and they were confirmed for the presence and expression of transgenes by the GUS assay, by the PCR and Southern blot and western blot analyses. Several T1 individuals from T0 line were evaluated for presence and expression of the nptII gene by the in vitro KR assay and by the PCR analysis. The presence of the HBsAg gene in the T1 individuals also was carried out by the PCR technique.
DOI: https://doi.org/10.31817/tckhnnvn.2014.12.7.
The effect of the Translation Inhibitor Cycloheximide on the Expression of AtZAT12and AtFITGenes in Arabidopsis

Le Thi Tuyet Cham, Vu Thi Thuy Hang

Date Received: 18-10-2022 / Date Accepted: 27-01-2023
Iron (Fe) is a microelement that plays an important role because it participates in redox reactions in plants. Fe uptake is tightly regulated to meet plant needs and is controlled by the transcription factor Fer-like iron deficiency-induced transcription factor (FIT). FIT expression increases in Fe deficiency but decreases in Fe deficient condition. However, when treated with Cycloheximide (CHX), this expression of FIT increased under Fe-sufficient conditions. This study aimed to evaluate whether ZAT12, an inhibitor of FIT, is associated with an elevation of FIT expression under conditions of Fe sufficiency and CHX treatment. The experiment evaluated the expression of FITand ZAT12after treatment with CHX at the end of treatment and 4 hours after treatment. The results showed that the expression of AtFIT and AtZAT12was elevated on Arabidopsis Col-0 and zat12-3grown under Fe sufficient conditions for 10 days and CHX treatment. Thus, in the absence of ZAT12 protein, either by mutations or by CHX translation inhibitors, AtFITexpression was increased. This study provides a way to remove FIT inhibition when plants grown in sufficient Fe condition.
Gene Expresssion of Cytokine and Toll-like Receptor Signaling molecule mRNA in Peripheral Blood Mononuclear Cells (PBMCs) from Pigs Co-infected with Porcine Reproductive and Respiratory Syndrome Virus and Porcine Circovirus

Dong Van Hieu, Phan Hong Dien, Tran Thi Huong Giang, Wen Bin Chung

Date Received: 18-03-2015 / Date Accepted: 18-05-2015
Field and experimental studies have shown that co-infection of PRRSV and PCV2 in pigs were quite common and thus the interactions between both viruses in the host might be critical to the pathogenesis of the disease. The exact interaction mechanisms are unknown and remained to be elucidated. In this study, the expression of cytokine mRNA as well as Toll like receptor signaling molecules associated with innate immunity in peripheral blood mononuclear cells (PBMCs) from pigs infected with PRRSV and/or PCV2 were investigated. Cytokine mRNA and TLR signaling molecule expression were determined by quantitative real-time polymerase chain reaction. Co-infection of pigs with PRRSV and PCV2 led to up-regulated expression of IL-1β and IL-10 mRNA and down-regulated expression of TNF-α and IL-6, IFN-α and IFN-β in PBMCs of pigs co-infected with PRRSV and PCV2. An increased expression of CD14 mRNA was noticed in pigs co-infected with PRRSV and PCV2. A down-regulated expression of IRF-1 and IRF-3 mRNA might also suppress the expression of IFNs that leads to the enhanced replication of PCV2 in pigs co-infected with PRRSV and PCV2.
Biểu hiện của lysozyme gen liên quan đến miễn dịch sau khi sử dụng chất kích thích miễn dịch trong tôm he Nhật Bản

Vu Duc Hanh, Nguyen Thi Hong Chien, Dao Le Anh, Nguyen Thi Hang, Le Van Truong, T. Itami, Lai Thi Lan Huong, Nguyen Thi Lan

Date Received: 26-07-2025 / Date Published: 06-08-2025
Chất kích thích miễn dịch là chất giúp tăng cường hoạt động của hàng rào miễn dịch và cung cấp sức đề kháng với bệnh. Ex400 được sản xuất từ vi tảo và Euglena là chất kích thích miễn dịch trên tôm được sử dụng để xác định biểu hiện gen của lysozyme trong tôm he Nhật Bản. ARN tổng số được chiết xuất từ các cơ quan lympho, ruột và máu của tôm tại các thời điểm 0, 3, 7 và 10 ngày sau khi cho tôm ăn Ex400. Trong ruột và cơ quan lympho của tôm, biểu hiện gen của lysozyme cao hơn so với lô đối chứng tại các thời điểm 7 và 10 ngày sau khi cho tôm ăn Ex400 (P< 0,01). Biểu hiện lysozyme trong máu của tôm cũng được xác định ở mức độ cao hơn so với lô đối chứng tại thời điểm 3 và 10 ngày sau khi cho ăn Ex400 (P < 0,01). Kết quả mức độ biểu hiện gen của lysozyme cho thấy chất kích thích miễn dịch có thể kích hoạt lysozyme trong hệ thống miễn dịch của tôm he Nhật Bản..
DOI: https://doi.org/10.31817/tckhnnvn.2017.15.3.
Investigating OsHSBP1 Gene Expression in Relation to Drought and Heat Responses in Rice (Oryza sativa L.)

Cao Le Quyen, Phung Thi Thu Huong, Dam Quang Hieu, Pham Xuan Hoi, Nguyen Duy Phuong

Date Received: 08-02-2025 / Date Published: 21-02-2025
The production of rice is significantly affected by adverse environmental conditions such as drought and heat. Heat shock proteins (HSPs) play a crucial role in mitigating stress tolerance responses associated with temperature and oxidation, thereby safeguarding cells and macromolecules from detrimental effects. The regulatory mechanism of HSP activity in some plant species involves the involvement of Heat Shock Factor Binding Protein (HSBP). This study aimed to investigate the functional activities and elucidate the stress response regulation mechanism of OsHSBP1 in rice. The expression pattern of this gene was analyzed under drought and heat stress conditions in two rice varieties, BC15 and OM54541. The OsHSBP1 expression increased at various time points during artificial heat and drought stress. The coding sequences of OsHSBP1 from these rice varieties were isolated and fully sequenced, revealing a gene segment of 237 base pairs in length. This segment encodes a polypeptide chain comprising 78 amino acids, including an α helix, exhibiting a similarity ranging from 59.8% to 97.4% homologous to HSBP proteins studied previously. These findings provide a foundation for exploring the functional role of OsHSBP1 to improve the drought and heat stress resistance of the major rice varieties such as BC15 and OM5451 through genetic engineering..
DOI: https://doi.org/10.1234/jq9q4727
Phân lập và khảo sát sự biểu hiện của promoter của gen phytase từ bacillus licheniformis DSM13

Nguyen Thanh Trung, Nguyen Minh Trung, Nguyen Huy Thuan, Trinh Thanh Trung, Nguyen Quoc Trung, Trinh Thi Thu Thuy, Nguyen Hoang Anh

Date Received: 20-09-2017 / Date Accepted: 25-03-2017 / Date Published: 06-08-2025
Phytase là một họ enzyme thủy phân cơ chất phytate để giải phóng ra phosphate vô cơ cùng các sản phẩm trung gian myo-inositol phosphate. Gene mã hóa enzyme phytase (phyL) ở Bacillus licheniformis DSM13 cảm ứng biểu hiện mạnh trong các điều kiện thiếu hụt phosphate. Trong nghiên cứu này, chúng tôi thiết kế vector biểu hiện sử dụng promoter của gene phyL để điều khiển sự biểu hiện của gen xynA (mã hóa enzyme xylanase). Hoạt tính promoter phyL được xác định thông qua mức độ biểu hiện của protein XynA. Chủng tái tổ hợp được nuôi cấy trong điều kiện môi trường không có hoặc có bổ sung phytate ở các nồng độ khác nhau. Kết quả cho thấy hoạt động của promoter phyL được cảm ứng mạnh khi nồng độ cơ chất phytate bổ sung vào ở giá trị £ 5 mM. Ngoài ra, thí nghiệm phân tích sinh trưởng của vi khuẩn trong môi trường có bổ sung phytate chỉ ra rằng phytate là một nguồn phosphate thay thế quan trọng giúp cho vi khuẩn B. licheniformis vượt qua các điều kiện đói phosphate.
DOI: https://doi.org/10.31817/tckhnnvn.2017.15.3.
Classifying Genes Encode Transmembrane Proteins Using Neighboring Genes

Tran Vu Ha, Pham Quang Dung, Nguyen Thi Thao, Doan Thi Thu Ha

Date Received: 06-10-2014 / Date Accepted: 20-12-2014
Like bio-diversity, there are too many proteins to experimentally annotate. Thus, methods for predicting the functions of proteins become necessary. In this article, we proposed a method that uses biological data to classify membrane transporters according to transported substrates. Motivated by the concept of Operons, our method used expression data and GO terms of neighboring genes to create input data for support vector machine. To rapidly obtain the result, we integrated LIBSVM in our tool then used this tool to train and test our classifiers. With this tool, users can classify membrane transporters and other kinds of proteins. This tool also allows users to add their desired organisms beside our tested ones.
Sự biểu hiện và di truyền các tính trạng ở quần thể F2 đậu xanh

Vu Thi Han, Vu Thi Thuy Hang, Vu Dinh Hoa

Date Received: 20-02-2017 / Date Accepted: 27-04-2017 / Date Published: 06-08-2025
Quần thể F2 của tổ hợp lai giữa Berken và ĐX92-1 và bố mẹ được trồng trên đồng ruộng, đánh giá và nghiên cứu sự biểu hiện và di truyền các tính trạng chất lượng và số lượng. Các tính trạng đánh giá đều phân ly. Các tính trạng chất lượng liên quan đến hình thái, quả và hạt được xác định do 1 hay 2 gen quy định, như màu sắc lá, hoa, hạt và rốn hạt, và độ bóng của hạt được do 1 gen điều khiển; hình dạng hạt, màu sắc quả, màu sắc thân do 2 gen điều khiển. Các đặc điểm về thời gian sinh trưởng không có sự biến động lớn ở F2. Tổng thời gian sinh trưởng của quần thể F2 dao động từ 71 - 76 ngày, chứng tỏ thời gian sinh trưởng ngắn. Giá trị trung bình đối với hầu hết các tính trạng số lượng về sinh trưởng, phát triển và năng suất cao hơn trung bình của bố mẹ. Một số cá thể có sự phân ly tăng tiến vượt ra ngoài khoảng của bố mẹ như kích thước lá, kích thước quả, năng suất cá thể, tổng khối lượng quả. CV% biến động từ 0.9 - 40.5%. Hệ số di truyền nghĩa rộng cho hầu hết các tính trạng thấp, 2.2 - 59.8%, cho thấy môi trường có ảnh hưởng lớn. Một số trường hợp đặc biệt bao gồm chiều rộng lá và quả có hệ số di truyền nghĩa rộng cao tương ứng với 90.4% và 97%; năng suất cá thể và khối lượng sinh khối khô có hệ số di truyền nghĩa rộng ở mức trung bình tương ứng với 64.9% và 66.8%. Năng suất cá thể trung bình của F2 tương đương với Berken. Một số cá thể có năng suất cao hơn Berken có thể được lựa chọn để nhân thế hệ và đánh giá tiếp.
DOI: https://doi.org/10.31817/tckhnnvn.2017.15.3.
Study on Expression of Chicken Interleukin 2 Recombinant Protein and its Modulate Cytokines Production in Macrophage Cell Line

Truong Anh Duc, Tran Thi Thanh Ha, Nguyen Thi Huyen, Chu Thi Nhu, Nguyen Thi Chinh, Hoang Van Tuan, Ly Duc Viet, Dang Vu Hoang, Yeong Ho Hong

Date Received: 20-11-2019 / Date Accepted: 16-06-2020
Interleukin-2 is a multifunctional cytokine, which modulates the proliferation and differentiation of various types of cells and induces cytokines expression. In this study, chicken IL-2 will be cloned, expressed and its functional characterization was evaluated in Macrophage (HD11) cell line. Genes coding for IL-2 (AF000631 from GenBank) linked with Trx-Tag thioredoxin leader and restriction sequences of EcoRI (5’end), HindIII (3’end) were cloned and then ligated into a cloning véc tơvector pCR2.1-TA. Chicken IL-2 gene was incorporated into pET32a(+) at restriction sites for the expression of the IL-2 gene in E. coliBL21. Chicken IL-2 recombinant protein produced in E. colisystem showed that it did not produce cytotoxicity and enhanced the proliferation of macrophage (HD11) cell line and induced IFNγ, IL-1β và and IL-6 mRNA expression. These results showed that recombinant chicken IL-2 protein production in E. colisystem was possessed natural biological properties and was recognized by specific antibodies against IL-2.
Biểu hiện gene mã hóa chitinase từ Bacillus licheniformis DSM13 trong E.coli T7 và xác định đặc tính của enzyme tái tổ hợp

Nguyen Hoang Anh

Date Received: 26-07-2017 / Date Accepted: 05-09-2017 / Date Published: 06-08-2025
Trong nghiên cứu này, gene mã hóa cho enzyme endochitinase của vi khuẩn gram dương Bacillus licheniformis DSM13 (ATCC 14580) được chuyển vào vectơ biểu hiện và biểu hiện trong tế bào E. coli T7. Mức độ biểu hiện cao của enzyme được thể hiện qua SDS-PAGE và hoạt tính của enzyme khi sử dụng pNP-(GlcNAc)2 0,18 mM là cơ chất. Hoạt tính của enzyme được biểu hiện trong E.coli T7 là cao khoảng gấp 8 lần so với enzyme biểu hiện ở Lactobacillus plantarum (nghiên cứu trước). Chitinase tái tổ hợp được tinh sạch và xác định đặc tính bằng cách sử dụng chitin 2% là cơ chất. Kết quả chỉ ra rằng, enzyme tái tổ hợp bền nhiệt (enzyme còn lại một nửa hoạt tính sau 15 và 8 ngày ủ ở 37 và 50°C tương ứng), bền ở giải pH từ 5 - 9. Sản phẩm chính thủy phân chitin huyền phù được xác định bằng phương pháp sắc ký bản mỏng là diacetyl glucucosamine. Các kết quả nghiên cứu này chỉ ra rằng enzyme tái tổ hợp này có tiềm năng trong việc ứng dụng sản xuất chitin-oligosaccharide, như là diacetyl glucucosamine chức năng từ việc chuyển hóa phế phụ phẩm chitin.
DOI: https://doi.org/10.31817/tckhnnvn.2017.15.9.
Establishment of ASimple and Efficient Transformation Protocol for Hairy Root Induction of Wild Bitter Melon (Momordica charantia L.var. abbreviata Ser.)

Nguyen Thi Thu Trang, Vu Thi Dao, Bui Minh Tri

Date Received: 28-09-2016 / Date Accepted: 13-02-2017
Wild bitter melon (Momordica charantia L. var. abbreviata Ser.) is known as a valuable vegetable and herb. It contains several important bioactive substances, including glycoside, saponin, alkaloide, triterpen and steroide. This study was carried out to establish a simple and efficient transformation protocol for inducting hairy roots of wild bitter melon by using strain TR7 of Agrobacterium rhizogenes which carries pPE113 binary vector harbouring e-gfp gene. Major parameters were optimized including, the age of plant and position for injection. The results indicated that highest hairy root induction frequencies was about 69.66-73.33% when seven-day-old seedlings were used to infect with the bacterium using hypocotyl. After regeneration and selection, thirty putative root clones of 4 specific phenotypes, i.e. white hairs, fast growth and branching on hormone-free MS medium, were obtained. The average GFP expression rate in 4 specific phenotypic groups were about 46.7%. PCR analysis using specific primers for rolA gene and virD gene confirmed the presence of the rolA gene and absence of virD gene in the genome of four hairy root lines. It was suggested that this simple transformation protocol can be applied for hairy root induction of other varieties as well.
Evaluation of AtZAT12Gene Expression in Transgenic Arabidopsis

Le Thi Tuyet Cham, Vu Ngoc Thang, Vu Thi Thuy Hang, Tran Anh Tuan

Date Received: 26-05-2020 / Date Accepted: 20-07-2020
ZAT12 is one of the transcription factors response to abiotic stress, which has had higher expression of ZAT12 under oxidative, cold, and heat stressetc. ZAT12 links iron deficiency and oxidative stress responses through direct interaction with and negative regulation of FIT.However, in thisstudy,ZAT12 was regulated by ZAT12native promoter. This study aimed to generate ZAT12 with overexpression (ZAT12 Ox) in response to oxidative stress and Fe uptake. The attempts of this study were further selected thetransgene lines containing 2x35S::ZAT12 by observing the dry seed under fluorescent microscopegene sequencing and PCR. The growth and gene expression as well asZAT12 protein regulation of 2x35S::ZAT12 lines (ZAT12 Ox)were evaluated. As the results showed, the 4 positive lines(CH1-1, 1-4, 1-10 and1-14lines) with different growth were generated, of which the growth of CH1-1 line was inhibited. Expression of AtZAT12 gene induced to response to Fe deficiency, but not to H2O2supplemented treatment. ZAT12 protein of CH1-4 line also detected by Western Blot.
Characterisation and Expression Survey of Dehydrin Gene Family in Clementine (Citrus clementina)

Cao Phi Bang

Date Received: 25-07-2014 / Date Accepted: 13-10-2014 / Date Published: 06-08-2025
A total of six dehydrin (DHN) genes have been found in the whole genome of Clementine (Citrus clementina). This work aimed to study the characteristics and structure of DHN in this species. All of six DHN proteins were highly hydrophilic. Their high aliphatic index suggested that clementine DHN might be thermostable proteins. Only two KS-type genes, CclDHN1 and CclDHN6, had no intron. Therefore, four others were discontinuous coding genes with an intron which separates two exons within the S fragment. The predicted 3D modeling structure showed many amphipathic α-helices in the secondary structure of DHN proteins. In addition, the serine phosphorylation sites also were predicted in the S fragment region. Two clementine DHN genes, CclDHN1 and CclDHN2, expressed in many different tissues under various environmental conditions .
DOI: https://doi.org/10.31817/tckhnnvn.2014.12.7.
Biểu hiện của gen ALF liên quan đến miễn dịch sau khi sử dụng chất kích thích miễn dịch trong tôm he Nhật Bản

Vu Duc Hanh, Lai Thi Lan Huong

Date Received: 31-07-2025 / Date Published: 06-08-2025
Chất kích thích miễn dịch là chất giúp tăng cường hoạt động của hàng rào miễn dịch và tác động đến các gen liên quan miễn dịch để bảo vệ của vật chủ. Ex400 được sản xuất từ vi tảo và Euglena là chất kích thích miễn dịch trên tôm được sử dụng để xác định biểu hiện của gen ALF trong tôm he Nhật Bản. ARN tổng số được chiết xuất từ các cơ quan lympho, ruột và máu của tôm tại các thời điểm 0, 3, 7 và 10 ngày sau khi cho tôm ăn Ex400. Trong ruột và cơ quan lympho của tôm, biểu hiện của gen ALF cao hơn so với lô đối chứng tại các thời điểm 3 ngày sau khi cho tôm ăn Ex400 (P< 0.01). Kết quả mức độ biểu hiện của gen ALF cho thấy chất kích thích miễn dịch có thể kích hoạt ALF trong hàng rào miễn dịch trong ruột và cơ quan lympho của tôm he Nhật Bản..
DOI: https://doi.org/10.31817/tckhnnvn.2017.15.9.
Co-expression of Human Interleukin-2 Gene with Chaperone GroESL and Thioredoxin in Escherichia coli BL21 (DE3)

Le Thi Lan Anh, Le Phuong Hoang Anh, Dao Trong Khoa, Le Thi Thu Hong, Phung Thu Nguyet, Nguyen Huu Duc, Nguyen Thi Hang, Truong Nam Hai

Date Received: 26-04-2013 / Date Accepted: 25-09-2013 / Date Published: 13-06-2025
Interleukin 2 (IL-2), one of the cytokines plays an important role for the immune response. IL-2 promotes the activation of T cells and so often used in immunotherapy such as kidney epithelial malignant and melanoma cancer. Gene encoding il-2 modified Cys125 to Ser was inserted into pET22b(+) harboring pelB secrete signal for expression in E. coli BL21(DE3) cells. To enhance the correct protein folding and solubility, IL-2 was simultaneously expressed with chaperone GroESL and thioredoxin (Trx). The results indicated that IL-2 proteins co-expressed with GroESL and Trx of 20 kDa were still existed in insoluble form and consisted of the pelB secret signal.
DOI: https://doi.org/10.31817/tckhnnvn.2013.11.6.
Genetic Association between Single Nucleotide Polymorphisms of PIT-1 Gene and Milk Production in Holstein Friesian Cows

Nguyen Thi Thu, Nguyen Thi Minh Hang, Nguyen Huu Duc, Ha Thanh Tùng, Le Thi Chau, Le Van Ty, Nguyen Thi Dieu Thuy

Date Received: 17-05-2013 / Date Accepted: 25-08-2013 / Date Published: 13-06-2025
PIT-1, a regulatory transcription factor, plays a role in pituitary cell differentiation and proliferation and expression level of hormones in the anterior pituitary gland. The objective of present study was to identify the single nucleotide polymorphisms of PIT-1 gene and their association with milk yield in Holstein Friesian (HF) cows. Milk yield was recorded for the whole lactation period. PCR-RFLP technique was applied for genotyping of two SNPs belonging to exon 2 (E2) and exon 6 (E6) with restriction enzymes TaqI and HinfI. The analysis of 53 Holstein milking cows showed that genotypic frequencies of AA, AB and BB of PIT-1E2 were 45.3, 28.3 and 26.4%, respectively. For the PIT-1E6, the highest frequency of GG genotype (73.6%), the lowest frequency of genotype TT (7.5%) and TG genotype (18.9%) were found. Minitab software was used to analyze genotype - phenotype association. The result revealed that theere exists positive association between locus PIT-1E2 with milk yield (P= 0.037 and ρ = 0.28) and number of milking day (P=0.015 and r= 0.306), whereas, no association of PIT-1E6 was found (P>0.05). Cows with AA genotype yielded more milk than those with BB and AB genotypes. These finding showed the potential use of AA genotype of PIT-1E2 in assisting breeding for increased milk production of Holstein Friesian cows.
DOI: https://doi.org/10.31817/tckhnnvn.2013.11.6.
The Crosstalk Between nicotine and PACAP in the Appetite Regulation in the Mouse Hypothalamus

Nguyen Thanh Trung, Yuki Kambe, Tran Thi Anh, Nguyen Manh Cuong

Date Received: 31-10-2022 / Date Accepted: 27-01-2023 / Date Published: 03-07-2025
Nicotine is one of the causes of suppressed appetite. Many reports have shown the underlying neurobiological mechanisms of the anorexia effects of cigarette smoking through nicotine receptors in the brain. However, whether nicotine may act through an additional pathway regulated by polypeptide pituitary adenylate cyclase (PACAP) in the brain is yet to be determined. To address these issues, we used pharmacological combination such as overexpression, knockdown PACAP gene in VMH and feeding in this study. At first, we found that dose-dependent nicotine decreased the food intake in mice treated daily for 5 days. In addition, more pronounced effects of inhibition feeding were observed in PACAP knockout mice. Furthermore, overexpression of PACAP in the VMH increased the expression while knockdown of PACAP in the VMH decreased the expression of α4 of Nicotine. Taken together, these results suggested that the expression of α4 receptor of Nicotine in the hypothalamus might be regulated by PACAP in the VMH.
DOI: https://doi.org/10.31817/tckhnnvn.2023.21.1.
Designing CRISPR/CAS9 System for Knocking out OsNRAMP5Related to Cadmium Accumulation in Rice Variety TBR225

Ngo Thi Van Anh, Nguyen Anh Minh, Pham Thu Hang, Pham Phuong Ngoc, Le Quynh Mai, Pham Xuan Hoi, Nguyen Duy Phuong

Date Received: 06-08-2022 / Date Accepted: 27-09-2022
Cadmium (Cd) is highly toxic to most living organisms, including humans. Rice with excessive Cd (> 0,2 mg/kg - QCVN 8-2:2011/BYT) is one of the leading causes of Cd poisoning, threatening human health. Several genes belonging to the NRAMP(natural resistance-associated macrophage protein) family are related to metal accumulation in plants; in which OsNRAMP5has been identified to be involved in the transport of various metals such as iron (Fe), zinc (Zn), copper (Cu), manganese) Mn and Cd in rice. In order to identify the function of OsNRAMP5in elite rice variety TBR225, we evaluated the expression of the targeted gene and designed Clustered regular interspaced short palindromic repeats/CRISPR-associated Cas9 (CRISPR/Cas9) system for editing TBR225OsNRAMP5. The increase of OsNRAMP5expression was observed in the root tissues of TBR225 rice plant in the Cd-treatment experiment using hydroponic culture. A sequence containing the ExonI region of OsNRAMP5with a size of 0.29kb was isolated from the total DNA of TBR225 to design single guide RNAs (sgRNA) for OsNRAMP5-editingCRISPR/Cas9 system. The sgRNA expression construct carrying two CRISPR RNA (crRNA) sequences recognizing two different sites on the ExonI of OsNRAMP5was ligated into the binary vector pCas9. The recombinant vector was validatedby PCR and sequencinganalysis. The results serve as basis for the improvement ofTBR225 rice variety with high quality byusinggenome editing technology.
Genetic Identification of Target Mutationsin OsSWEET14 Promoter-edited TBR225 Rice Lines

Tran Lan Dai, Pham Thu Hang, Cao Le Quyen, Pham Thi Van, Pham Xuan Hoi, Nguyen Duy Phuong

Date Received: 29-11-2021 / Date Accepted: 05-04-2022
Understanding host-pathogen interactions is key in breeding disease-resistant crops, including bacterial leaf blight. As a key virulence strategy to cause bacterial leaf blight, Xanthomonas oryzae pv.oryzae(Xoo) secretes transcription activator-like effectors (TALEs) to control the expression of host target genes through binding to the effector-binding elements (EBEs) located on the promoter regions. Precise mutagenesis at the TALE-interaction sites on the host genes by using gene editing tools such as clustered regularly interspaced short palindromic repeats/CRISPR-associated protein-9 nuclease (CRSIPR/Cas9) has been seen as a promising way to improve BLB resistance ofrice cultivars. In a previous study, we generated some transgenic TBR225 rice lines containing a single copy of CRISPR/Cas9 expression construct for editing AvrXa7/Tal5EBE on the promoter OsSWEET14. In this study, we continued to evaluate genotype and phenotype of T1edited TBR225 lines to determinethe genetic stability and phenotypic effective of the CRISPR/Cas9-induced OsSWEET14mutations. The genotypic analysis showed that 65%of transgenic TBR225 plants harbored the targeted OsSWEET14mutation, including nucleotide (Nu) insertion and deletion. The OsSWEET14mutations were stably inherited to the next generation with an expected segregation ratio consistent with Mendelian segregation (1:2:1). The preliminary phenotypic analysis suggested that the mutation of OsSWEET14promoter did not affect the growth and yield of gene-edited rice plants. Some T1TBR225 lines showed the resistance to Xoo isolate VXO_11 in the leaf-clipping experiment. The obtained results are a premise for development of the BLB-resistant TBR225 rice variety in the future.
Insect ResistanceGene(GNA)Transformationinto Cottonby Agrobacterium tumefaciens

Pham Duc Tri, Hoang Van Duong, Le Tan Duc, Phan Tuong Loc, Mai Truong, Nguyen Huu Ho

Date Received: 05-01-2015 / Date Accepted: 22-07-2015
Hypocotyl explants of cotton (Gossypium hirsutum L.) cv. Coker 312 were transformed with Agrobacterium tumefaciens strain EHA 105 containing the GNA gene under the control of RSs1 promoter and selected on callus initiation medium containing kanamycin. Somatic embryogenesis was promoted in the surviving calli. Plantlets were recovered from putative transgenic embryos. Polymerase Chain Reaction (PCR) and Southern blot analysis were used to confirm the integration of GNA transgene into the plant genome. Western blot result indicated the expression of the GNA gene in transgenic cotton plants.

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