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编码氯霉素乙酰转移酶的大肠杆菌质粒基因在酵母中的功能性表达。

Functional expression in yeast of the Escherichia coli plasmid gene coding for chloramphenicol acetyltransferase.

作者信息

Cohen J D, Eccleshall T R, Needleman R B, Federoff H, Buchferer B A, Marmur J

出版信息

Proc Natl Acad Sci U S A. 1980 Feb;77(2):1078-82. doi: 10.1073/pnas.77.2.1078.

Abstract

The Escherichia coli R factor-derived chloramphenicol resistance (camr) gene is functionally expressed in the yeast Saccharomyces cerevisiae. the gene was introduced by transformation into yeast cells as part of a chimeric plasmid, pYT11-LEU2, constructed in vitro. The plasmide vector consists of the E. coli plasmid pBR325 (carrying the camr gene), the yeast 2-micron DNA plasmid, and the yeast LEU2 structural gene. Yeast cells harboring pYT11-LEU2 acquire resistance to chloramphenicol and cell-free extracts prepared from such cells contain chloramphenicol acetyltransferase (acetyl-CoA: chloramphenicol 3-O-acetyltransferase, EC 2.3.1.28), the enzyme specified by the camr gene in E. coli. Resistance to chloramphenicol and the presence of chloramphenicol acetyltransferase activity segregate with the yeast marker LEU2, carried by the transforming plasmid, during both mitotic growth and meiotic division.

摘要

源自大肠杆菌R因子的氯霉素抗性(camr)基因在酿酒酵母中实现了功能性表达。该基因作为体外构建的嵌合质粒pYT11-LEU2的一部分,通过转化导入酵母细胞。质粒载体由大肠杆菌质粒pBR325(携带camr基因)、酵母2-μm DNA质粒和酵母LEU2结构基因组成。携带pYT11-LEU2的酵母细胞获得了对氯霉素的抗性,并且从此类细胞制备的无细胞提取物含有氯霉素乙酰转移酶(乙酰辅酶A:氯霉素3-O-乙酰转移酶,EC 2.3.1.28),该酶由大肠杆菌中的camr基因所编码。在有丝分裂生长和减数分裂过程中,对氯霉素的抗性以及氯霉素乙酰转移酶活性的存在与转化质粒携带的酵母标记LEU2一起分离。

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