梅花鹿大肠杆菌的分离与鉴定

Isolation and Identification of Escherichia coli from Dead Sika

本试验采集武威市天祝县某养殖场病死梅花鹿病料,经细菌分离培养、细菌形态观察、培养特性、生化特性鉴定、药敏试验及致病性检测,初步确定致病菌种。对已分离经纯培养的菌株提取基因组DNA,利用引物进行PCR扩增,并得到预期的基因片段。切下目的条带分别进行回收、载体与产物的连接转化,最后挑取阳性斑置于LB肉汤中,送生物公司测序后,结合细菌学试验结果最终确定该梅花鹿死亡是由大肠杆菌引起,在此基础上提出综合性的防治措施。

Pathological samples of dead sika was collected from a farm of Tianzhu,pathogenic bacterium was identified primitively by culturing isolated bacterial,observing morphology,identifying biological trait,medical sensitivity experiment and animal pathogenicity test.We extracted genomic DNA of the isolated strain,used primers for PCR amplification and got the expected gene fragment.Cutting the target band and proceeding gel extraction,production and pGEM-T was ligated and transformed.Finally,positive blot was picked and cultured in LB-Broth-Medium.Combined the result of bacteriological experiment and the result of biology company sequenced,Escherichia coli was identified finally,and there are some omnibus control measures was adopted on the basis of result.