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海洋硅藻圆柱鞘藻属N1菌株叶绿体编码的rbcL和rbcS基因的克隆与表达

Cloning and expression of the chloroplast-encoded rbcL and rbcS genes from the marine diatom Cylindrotheca sp. strain N1.

作者信息

Hwang S R, Tabita F R

机构信息

Department of Microbiology, Ohio State University, Columbus 43210.

出版信息

Plant Mol Biol. 1989 Jul;13(1):69-79. doi: 10.1007/BF00027336.

Abstract

Both the rbcL and rbcS genes, encoding the large and small subunits, respectively, of ribulose 1,5-bisphosphate carboxylase/oxygenase, have been found to be encoded by chloroplast DNA in the marine diatom Cylindrotheca sp. N1. The rbcS gene in this diatom was found to be adjacent to the rbcL gene by a combination of: (i) Southern-blotting analyses, using heterologous probes; (ii) examination of recombinant proteins synthesized in Escherichia coli, directed by cloned rbcL/rbcS genes; and (iii) synthesis of enzymatically active heterologous Rubisco protein in vivo by recombinant DNA procedures using large subunits of Anacystis nidulans and small subunits of Cylindrotheca sp. N1. It appears that two copies of rbcL and rbcS genes are encoded by the chloroplast DNA of this diatom.

摘要

在海洋硅藻圆柱鞘丝藻(Cylindrotheca sp. N1)中,分别编码1,5-二磷酸核酮糖羧化酶/加氧酶大亚基和小亚基的rbcL基因和rbcS基因,均已被发现由叶绿体DNA编码。通过以下方法相结合,发现该硅藻中的rbcS基因与rbcL基因相邻:(i)使用异源探针进行Southern杂交分析;(ii)检测由克隆的rbcL/rbcS基因指导在大肠杆菌中合成的重组蛋白;(iii)通过使用集胞藻6803的大亚基和圆柱鞘丝藻N1的小亚基,采用重组DNA程序在体内合成具有酶活性的异源Rubisco蛋白。该硅藻的叶绿体DNA似乎编码了两个拷贝的rbcL基因和rbcS基因。

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