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 μg 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.

中文翻译：

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pGP-B2E，一种重组兼容的TA / TB连接载体，用于快速廉价的基因克隆。

DNA克隆是生命科学各个领域的基本步骤，并且已经进行了许多努力来简化此过程。在这项研究中，我们报告了两个通用质粒（pGP），pGP-XB2E和pGP-B2E，用于快速，经济高效地构建基本克隆。在pGP-XB2E中设计了BciVI和XcmI切割位点，以测试质粒的线性化效率。通过使用BciVI，该质粒具有更好的线性化效率，该BciVI可以在10分钟内几乎完全消化2μg质粒，而推荐酶浓度仅为十分之一。为了进一步优化pGP-XB2E，设计了去除XcmI切割位点的新质粒pGP-B2E。该载体可高效克隆高达1812 bp的PCR产物，菌落数约为pGP-XB2E的5倍。除了TA克隆外，T末端为引物的平末端PCR产物可与T载体pGP-B2E正相关，而无需A-尾处理（TB克隆）。此外，作为进入载体，pGP-B2E也可用于无移码突变的网关重组反应。通常，此载体为基础分子克隆提供了一种通用，快速且经济高效的方法。