pGP-B2E, a Recombinant Compatible TA/TB-Ligation Vector for Rapid and Inexpensive Gene Cloning
文献类型： 外文期刊
作者： Li, Dongyue 1 ; Zheng, Chao 2 ; Zhou, Jie 2 ; Chen, Bin 2 ; Xu, Rumeng 2 ; Yuan, Wenxia 2 ; Zheng, Ersong 2 ; Liang, Weif 1 ;
作者机构： 1.Yunnan Agr Univ, Coll Plant Protect, Kunming 650201, Yunnan, Peoples R China
2.Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Agr Prod Qua, MOA Key Lab Plant Protect & Biotechnol, Zhejiang Prov Key Lab Plant Virol, Hangzhou 310021, Zhejiang, Peoples R China
3.Northwest A&F Univ, Coll Plant Protect, Yangling 712100, Shaanxi, Peoples R China
4.Zhejiang Normal Univ, Coll Chem & Life Sci, Jinhua 321004, Zhejiang, Peoples R China
5.Ningbo Univ, Inst Plant Virol, Ningbo, Zhejiang, Peoples R China
6.Ningbo Acad Agr Sci, Inst Biotechnol, Ningbo 315101, Zhejiang, Peoples R China
关键词： TA cloning; Blunt-end ligation; Gateway entry vector; High efficiency; Low cost
期刊名称：MOLECULAR BIOTECHNOLOGY （ 影响因子：2.695； 五年影响因子：2.303 ）
ISSN： 1073-6085
年卷期： 2020 年 62 卷 1 期
页码：
收录情况： SCI
摘要： DNA cloning is the basic step for different fields of life science, and many efforts have been made to simplify this procedure. In this study, we report two general purpose plasmids (pGP), pGP-XB2E and pGP-B2E, for rapid and cost-effective construct of basic clones. The BciVI and XcmI cleavage sites are designed in pGP-XB2E to test plasmid linearization efficiency. The plasmid has better linearization efficiency by using BciVI which could almost completely digest 2 mu g plasmid in 10 min with only one-tenth the recommended enzyme concentration. In order to further optimize the pGP-XB2E, a new plasmid, pGP-B2E, which removed XcmI cleavage site was designed. This vector is highly efficient for cloning PCR products up to 1812 bp, and the number of colonies was about five times that of pGP-XB2E. In addition to TA cloning, blunt-end PCR products with T ended in the primer could be positively linked to the T-vector pGP-B2E without A-tailing treatment (TB cloning). Moreover, as an entry vector, pGP-B2E was also compatible for gateway recombination reaction without frameshift mutations. In general, this vector provides a universal, quick, and cost-efficient method for basic molecular cloning.
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