嗜水气单胞菌胞外蛋白酶编码基因（eprA1）的克隆与序列分析

Cloning and sequence analysis of the gene (eprA1) encoding an extracellular protease from Aeromonas hydrophila.

作者信息

Chang T M, Liu C C, Chang M C

机构信息

Department of Biochemistry, Medical College, National Cheng Kung University, Tainan, Taiwan.

出版信息

Gene. 1997 Oct 15;199(1-2):225-9. doi: 10.1016/s0378-1119(97)00371-5.

DOI:10.1016/s0378-1119(97)00371-5

PMID:9358060

Abstract

A gene (eprA1) encoding the extracellular protease of Aeromonas hydrophila AH1 has been cloned and sequenced. Nucleotide sequence analysis of eprA1 predicted a single open reading frame (ORF) of 1038 bp encoding a 346 amino acid (aa) polypeptide, with a potential 21-aa signal peptide. When the eprA1 gene was expressed in minicells, one major band of approx. 37 kDa was identified, while protease activity staining experiments identified a caseinolytic band of approx. 29 kDa determined by SDS-PAGE analysis of the minicells. The deduced C-terminal aa region (Arg-290 to Gly-313) showed sequence homology to partial C-terminal sequences of other zinc metalloproteases including Penicillium citrinum metalloprotease (PlnC), Aspergillus oryzae metalloprotease (NpII), Aspergillus flavus metalloprotease (MepA), and Aspergillus fumigatus metalloprotease (Mep20), particularly with respect to zinc-binding residues.

摘要

编码嗜水气单胞菌AH1胞外蛋白酶的基因（eprA1）已被克隆并测序。eprA1的核苷酸序列分析预测有一个1038 bp的单一开放阅读框（ORF），编码一个346个氨基酸（aa）的多肽，带有一个潜在的21个氨基酸的信号肽。当eprA1基因在微细胞中表达时，鉴定出一条约37 kDa的主要条带，而蛋白酶活性染色实验通过对微细胞的SDS-PAGE分析鉴定出一条约29 kDa的酪蛋白水解条带。推导的C末端氨基酸区域（Arg-290至Gly-313）与其他锌金属蛋白酶的部分C末端序列具有序列同源性，包括桔青霉金属蛋白酶（PlnC）、米曲霉金属蛋白酶（NpII）、黄曲霉金属蛋白酶（MepA）和烟曲霉金属蛋白酶（Mep20），特别是在锌结合残基方面。