三株弧菌热休克蛋白70基因的克隆和序列分析

 根据GenBank中同类蛋白序列设计特异PCR引物，从2株创伤弧菌Vibrio vulnificus和1株河弧菌Vibrio fluvialis中扩增出热休克蛋 白70（heat shock protein, hsp70）基因片段。对这3个片段进行克隆、测序和分析的结果表明，3个片段长均为1 911 bp，包含完整的 hsp70 ORF，编码636个氨基酸。它们的氨基酸序列与GenBank中其它物种hsp70的氨基酸序列比较发现，2株创伤弧菌hsp70基因序列 和同种其它菌株的同源性高，达98%以上；而河弧菌的hsp70序列属首次克隆；与多种原核和真核生物的hsp70氨基酸序列一起构建 了系统进化树，结果支持传统的分类结果。

Cloning and Sequence Analysis of hsp70 of Three Strains of Vibrio

A pair of primers was designed according to the nucleotide sequence of the heat shock protein (hsp70) gene of vibrio reported in GenBank.With the specific primers, three target fragments were amplified from two of Vibrio vulnificus strains and a Vibrio fluvialis stain genomic DNA by PCR.The fragments were cloned and sequenced.The result showed that they included an ORF of 1911 bp which encoded a polypeptide of 636 amino acids.The identities of hsp70 among these two Vibrio vulnificus stains and other Vibrio vulnificus stains from GenBank are more than 98%.The hsp70 from Vibrio fluvialis was cloned for the first time.The neighbor joining tree of hsp70 was constructed and it supported the traditional classification.