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对编码集胞藻6803光合系统I一个亚基的psaI基因进行靶向失活。

Targeted inactivation of the gene psaI encoding a subunit of photosystem I of the cyanobacterium Synechocystis sp. PCC 6803.

作者信息

Nakamoto H

机构信息

Department of Biochemistry and Molecular Biology, Faculty of Science, Saitama University, Urawa, Japan.

出版信息

Plant Cell Physiol. 1995 Dec;36(8):1579-87.

PMID:8589933
Abstract

The psaI gene encoding the 4 kDa polypeptide subunit (PsaI) of the photosystem I was cloned from the Synechocystis sp. PCC 6803, a readily transformable cyanobacterium. The gene is located immediately downstream of the psaL gene in this cyanobacterial genome and encodes a polypeptide of 40 amino acid residues with a deduced molecular mass of 4,413 Da. The deduced amino acid sequence is homologous to PsaI from other cyanobacterial and higher plants. The poly-peptide has an internal hydrophobic region with a hydrophilic C-terminal region. The gene is present as a single copy in the genome. While some of its transcripts are expressed as polycistronic mRNAs which are co-transcribed with psaL, the other may be expressed as monocistronic mRNAs. Stable mutant strains in which the gene psaI was interrupted by a gene conferring resistance to kanamycin, were generated by targeted mutagenesis. One of the mutants was characterized in detail. SDS polyacrylamide gel electrophoresis revealed that the mutant strain contained other polypeptides of photosystem I in thylakoid membranes. Growth and photosynthetic characteristics of the mutant strain under photoautotrophic conditions were similar to those of the wild type, suggesting a dispensable unknown role of PsaI for photosynthesis in Synechocystis sp. PCC 6803.

摘要

从易于转化的蓝藻集胞藻PCC 6803中克隆了编码光系统I的4 kDa多肽亚基(PsaI)的psaI基因。该基因位于此蓝藻基因组中psaL基因的紧邻下游,编码一个由40个氨基酸残基组成的多肽,推导分子量为4413 Da。推导的氨基酸序列与其他蓝藻和高等植物的PsaI同源。该多肽有一个内部疏水区域和一个亲水的C末端区域。该基因在基因组中以单拷贝形式存在。其一些转录本以与psaL共转录的多顺反子mRNA形式表达,另一些可能以单顺反子mRNA形式表达。通过定点诱变产生了psaI基因被赋予卡那霉素抗性的基因中断的稳定突变株。对其中一个突变株进行了详细表征。SDS聚丙烯酰胺凝胶电泳显示,突变株在类囊体膜中含有光系统I的其他多肽。该突变株在光合自养条件下的生长和光合特性与野生型相似,表明PsaI在集胞藻PCC 6803的光合作用中具有一个可有可无的未知作用。

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