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分别编码来自鱼腥藻Anabaena sp. PCC 7119的质体蓝素和细胞色素c6的petE和petJ基因在大肠杆菌中的克隆及正确表达。

Cloning and correct expression in Escherichia coli of the petE and petJ genes respectively encoding plastocyanin and cytochrome c6 from the cyanobacterium Anabaena sp. PCC 7119.

作者信息

Molina-Heredia F P, Hervás M, Navarro J A, De la Rosa M A

机构信息

Instituto de Bioquímica Vegetal y Fotosíntesis, Centro Isla de la Cartuja, Universidad de Sevilla, Spain.

出版信息

Biochem Biophys Res Commun. 1998 Feb 4;243(1):302-6. doi: 10.1006/bbrc.1997.7953.

Abstract

The genes coding for plastocyanin (petE) and cytochrome c6 (petJ) from Anabaena sp. PCC 7119 have been cloned and properly expressed in Escherichia coli. The recombinant proteins are identical to those purified from the cyanobacterial cells. The products of both the petE and petJ genes are correctly processed in E. coli, as deduced from their identical N-terminal amino acid sequences as compared with those of the metalloproteins isolated from the cyanobacterium. Physicochemical and functional properties of the native and recombinant protein preparations are also identical, thereby confirming that expression of petE and petJ genes in E. coli is an adequate tool to address the study of the structure/function relationships in plastocyanin and cytochrome c6 from Anabaena by site-directed mutagenesis.

摘要

已从鱼腥藻属PCC 7119中克隆出编码质体蓝素(petE)和细胞色素c6(petJ)的基因,并在大肠杆菌中正确表达。重组蛋白与从蓝藻细胞中纯化得到的蛋白相同。根据petE和petJ基因产物与从蓝细菌中分离出的金属蛋白的N端氨基酸序列相同,推断它们在大肠杆菌中得到了正确加工。天然和重组蛋白质制剂的物理化学和功能特性也相同,从而证实了在大肠杆菌中表达petE和petJ基因是通过定点诱变研究鱼腥藻属质体蓝素和细胞色素c6结构/功能关系的合适工具。

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