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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
2013-03-29
发表期刊PLOS ONE
ISSN1932-6203
卷号8期号:3页码:e59576
摘要With 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.
关键词Protein-kinase Salt Tolerance Arabidopsis Versatile Cloning Expression System Genes Transformation Encodes
资助信息National Sciences Foundation of China [30971452, 31170256]; Major Sciences and Technology Program [110201101003-TS-03]; Young Academic and Technical Leader Raising Foundation of Yunnan Province [2012HB041]; State Education Ministry
学科领域Science & Technology - Other Topics
收录类别SCI
语种英语
WOS研究方向Science & Technology - Other Topics
WOS类目Multidisciplinary Sciences
WOS记录号WOS:000317263900018
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文献类型期刊论文
条目标识符http://ir.kib.ac.cn/handle/151853/16513
专题中国科学院青藏高原研究所昆明部
作者单位1.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
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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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