水貂铜绿假单胞菌鞭毛分型及分离株flic基因的克隆与序列分析

为分析铜绿假单胞菌(PA)分离株鞭毛蛋白基因(flic)之间的差异,本研究通过PCR方法扩增由山东地区发病水貂的肺脏组织中分离的29株分离株flic基因的部分片段,根据其片段长度确定PA的鞭毛类型。选取两种类型代表株(PASD01、PASD03),采用PCR方法扩增flic全基因进行序列分析。结果表明,PASD01分离株与国外A型PA flic基因序列同源性为99.6%～99.7%,PASD03分离株与B型PA flic基因的序列同源性为98.9%～99.5%,而两个分离株之间的序列同源性为76%。本研究首次从水貂病变组织中克隆得到两种鞭毛蛋白类型的flic基因,该基因在各型鞭毛菌株中高度保守,为进一步研制鞭毛亚单位疫苗提供实验依据。

Flagellar identification and sequence analysis of flic gene of Pseudomonas aeruginosa isolated from mink

To identify the flagellum type of Pseudomonas aeruginosa,the partial fragments of flic gene were amplified from 29 isolates of P.aeruginosa isolated in lung tissues of infected minks by PCR,According to the size of the PCR product,18 isolates were flagellum type A(1,164 bp or 1,185 bp) and 11 isolates were type B(1,467 bp),respectively.Furthermore,the complete flic genes of P.aeruginosa type A(PASD01) and type B(PASD03) were amplified and sequenced.The nucleotide sequence alignment showed that the flic gene of PASD01 shared 99.6% to 99.7% identity with serotype A reference strains and PASD03 shared 98.9% to 99.5% identity with serotype B strains,respectively.However,PASD01 was only 76% identity with PASD03.This was the first reported to identify 2 types of flic genes from diseased minks,which provided basis for the further development of the flagella subunit vaccine against P.aeruginosa infedtion in minks.