沙眼衣原体ompA真核表达重组体的构建及鉴定

目的 扩增D型沙眼衣原体ompA基因，构建真核表达重组质粒，为核酸疫苗的研制做准备。方法 用PCR技术从D型Ct基因组DNA中扩增ompA基因片段，重组入pUCm—T克隆载体。将pUCm—T／ompA中的ompA外源基因片段经酶切、连接等反应，亚克隆入pcDNA3．1( )真核表达载体，再经含氨苄青霉素的LB培养基筛选，酶切、PCR扩增及测序鉴定。结果 从D型ct基因组DNA中扩增出特异的ompA基因片段，筛选出pUCm—T／ompA；经亚克隆，筛选鉴定出pcDNA3．1( )／ompA重组质粒。结论 成功地构建了pUCm—T／ompA重组质粒和pcLDNA3．1( )／ompA重组质粒，为Ct核酸疫苗的研制提供实验基础。

Construction and Identification of Eukaryotic Expression OmpA Recombinant of Chlamydia Trachomatis

Objective To construct the recombinant plasmid of pcDNA3.1(+)/ompA. Methods OmpA gene was amplified from the genomic DNA of Chlamydia trachomatis using polymerase chain reaction (PCR),which was inserted into cloning vector,pUCm-T.The inserted ompA gene was subcloned to pcDNA3.1(+) eukaryotic expression vector by digesting with restrictive enzymes and linking reactions.The positive clone was screened on LB plates containing ampicillin and identified by restrictive enzymes digestion, PCR amplification and sequencing.Results The specific gene fragment ompA was amplified; pUCm-T/ompA and pcDNA3.1(+)/ompA were constructed.Conclusion pcDNA3.1(+)/ompA recombinant was successfully constructed.