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源自pACYC184的克隆载体,包含pUC8/9和pUC18/19质粒的多克隆位点和lacZα报告基因。

pACYC184-derived cloning vectors containing the multiple cloning site and lacZ alpha reporter gene of pUC8/9 and pUC18/19 plasmids.

作者信息

Martinez E, Bartolomé B, de la Cruz F

机构信息

Departamento de Biología Molecular, Universidad de Cantabria, Santander, Spain.

出版信息

Gene. 1988 Aug 15;68(1):159-62. doi: 10.1016/0378-1119(88)90608-7.

Abstract

A new series of vectors, pSU2716, pSU2717, pSU2718, and pSU2719, has been constructed. The plasmids contain (i) the P15A replicon, (ii) the chloramphenicol acetyl transferase (CAT)-coding gene from Tn9, and (iii) the HaeII fragment which carries the multiple cloning site and the lacZ alpha reporter gene of pUC8, pUC9, pUC18 and pUC19, respectively. These vectors allow rapid and simple transfer of inserts from pUC plasmids, have an intermediate copy number (which allows regulated expression from the lac promoter), and are compatible with ColE1-derived vectors (and, therefore, can be used in studies requiring the joint expression of two genes, for example, in genetic complementation analysis). Furthermore, the accumulation of CAT instead of beta-lactamase, allows an easy visualization in sodium dodecyl sulfate-polyacrylamide-gel electrophoresis of proteins of 28-35 kDa, which can otherwise be obscured by the beta-lactamase.

摘要

构建了一系列新的载体,即pSU2716、pSU2717、pSU2718和pSU2719。这些质粒包含:(i)P15A复制子;(ii)来自Tn9的氯霉素乙酰转移酶(CAT)编码基因;(iii)HaeII片段,其分别携带pUC8、pUC9、pUC18和pUC19的多克隆位点和lacZα报告基因。这些载体能快速、简单地从pUC质粒转移插入片段,具有中等拷贝数(可实现来自lac启动子的调控表达),并且与源自ColE1的载体兼容(因此可用于需要两个基因联合表达的研究,如基因互补分析)。此外,积累的是CAT而非β-内酰胺酶,这使得在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中能轻松观察到28 - 35 kDa的蛋白质,否则这些蛋白质会被β-内酰胺酶掩盖。

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