摘要
目的 构建金黄色葡萄球菌肠素D(SED)毒力蛋白基因重组表达质粒。方法 根据SED已知序列 ,设计合成一对引物 ,用PCR方法从金黄色葡萄球菌毒力质粒上扩增出基因片段 ,克隆到原核表达质粒PET3 2中 ,转化大肠埃希菌JM 10 9感受态细胞 ,经酶切和PCR鉴定 ,然后进行测序。结果 PCR扩增产物大小为 3 17bp ;重组质粒经双酶切和PCR鉴定表明已正确重组 ;测序结果与己知序列基本吻合。结论 国内首次克隆了金黄色葡萄球菌肠毒素D产毒基因 。
ObjectiveTo construct a recombinant plasmid containing s taphylococcus aureus toxic protein gene. MethodsA couple of primers were designed for PCR accordi ng to the known sequence of SED.The gene obtained by amplification from plasmid DNA of staphylococcus aureus by PCR technique was cloned into plamid of pET32a d irectionally.The recombinant plasmid pET32a was transferred into competent JM109 .The recombinants were screened and identified by restriction analysis and PCR,t he cloned gene was sequened.ResultsThe size of amplified PCR products was 317bp.The correct recombinant plasmid pET32a was isolated and confirmed by restriction ana lysis and PCR,DNA sequencing showed the DNA sequence of the cloned gene was the same as the published sequence.ConclusionsThe SED toxic gene was first successfully amp lified and cloned into plasmid PET32a in our country.It provided the basic mater ial for studying the pathogenesis.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2004年第3期274-275,共2页
Chinese Journal of Public Health
基金
全军医药卫生"十五"指令性课题 ( 0 1L 0 50 )
