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A Series of TA-Based and Zero-Background Vectors for Plant Functional Genomics
Wang, Chuntao1,2; Yin, Xianlun1,2; Kong, Xiangxiang1,3; Li, Wansha1,2; Ma, Lan1,3; Sun, Xudong1,2; Guan, Yanlong1,3; Todd, Christopher D.4; Yang, Yongping1,2; Hu, Xiangyang1,2
Corresponding AuthorHu, XY (reprint author), Chinese Acad Sci, Kunming Inst Bot, Key Lab Biodivers & Biogeog, Kunming, Yunna, Peoples R China. ; huxiangyang@mail.kib.ac.cn
2013-03-29
Source PublicationPLOS ONE
ISSN1932-6203
Volume8Issue:3Pages:e59576
AbstractWith the sequencing of genomes from many organisms now complete and the development of high-throughput sequencing, life science research has entered the functional post-genome era. Therefore, deciphering the function of genes and how they interact is in greater demand. To study an unknown gene, the basic methods are either overexpression or gene knockout by creating transgenic plants, and gene construction is usually the first step. Although traditional cloning techniques using restriction enzymes or a site-specific recombination system (Gateway or Clontech cloning technology) are highly useful for efficiently transferring DNA fragments into destination plasmids, the process is time consuming and expensive. To facilitate the procedure of gene construction, we designed a TA-based cloning system in which only one step was needed to subclone a DNA fragment into vectors. Such a cloning system was developed from the pGreen binary vector, which has a minimal size and facilitates construction manipulation, combined with the negative selection marker gene ccdB, which has the advantages of eliminating the self-ligation background and directly enabling high-efficiency TA cloning technology. We previously developed a set of transient and stable transformation vectors for constitutive gene expression, gene silencing, protein tagging, subcellular localization analysis and promoter activity detection. Our results show that such a system is highly efficient and serves as a high-throughput platform for transient or stable transformation in plants for functional genome research.
KeywordProtein-kinase Salt Tolerance Arabidopsis Versatile Cloning Expression System Genes Transformation Encodes
Subject AreaScience & Technology - Other Topics
Indexed BySCI
Language英语
WOS Research AreaScience & Technology - Other Topics
WOS SubjectMultidisciplinary Sciences
WOS IDWOS:000317263900018
Citation statistics
Cited Times:8[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.kib.ac.cn/handle/151853/16513
Collection中国科学院青藏高原研究所昆明部
Affiliation1.Chinese Acad Sci, Kunming Inst Bot, Key Lab Biodivers & Biogeog, Kunming, Yunna, Peoples R China
2.Chinese Acad Sci, Kunming Inst Bot, Plant Germplasm & Genom Ctr, Germplasm Bank Wild Species, Kunming, Yunna, Peoples R China
3.Univ Chinese Acad Sci, Beijing, Peoples R China
4.Univ Saskatchewan, Dept Biol, Saskatoon, SK S7N 0W0, Canada
Recommended Citation
GB/T 7714
Wang, Chuntao,Yin, Xianlun,Kong, Xiangxiang,et al. A Series of TA-Based and Zero-Background Vectors for Plant Functional Genomics[J]. PLOS ONE,2013,8(3):e59576.
APA Wang, Chuntao.,Yin, Xianlun.,Kong, Xiangxiang.,Li, Wansha.,Ma, Lan.,...&Hu, Xiangyang.(2013).A Series of TA-Based and Zero-Background Vectors for Plant Functional Genomics.PLOS ONE,8(3),e59576.
MLA Wang, Chuntao,et al."A Series of TA-Based and Zero-Background Vectors for Plant Functional Genomics".PLOS ONE 8.3(2013):e59576.
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