堆型艾美尔球虫cSZ1基因在PMV361整合载体的构建

Eimeria acervulina cSZ1 Gene Construction in PMV361 Vectors

目的:构建PMV361-cSZ1载体,研制鸡球虫疫苗。方法:根据堆型艾美尔球虫基因序列,设计合成引物,扩增cSZ1基因,在pMD18-T Simple Vector表达后提取质粒,经过PCR鉴定和酶切鉴定。将cSZ1基因片段和PMV361载体由限制性内切酶PvuⅡ和ClaI双酶切后,进行DNA连接。结果:经PT-PCR扩增出cSZ1基因片段,大小为940bp,连接到整合表达载体PMV361中,成功构建PMV361-cSZ1的整合表达载体。结论:成功扩增了堆型艾美耳球虫cSZ1基因开放阅读框并构建了整合表达载体pMV361-cSZ1,为构建鸡球虫重组卡介疫苗内表达提供前期实验基础。

To construct PMV361-cSZ1 Construction,development of chicken coccidiosis vaccine.Method according to Eimeria acervulina cSZ1 gene sequence,were designed and synthesized primers,the expression of pMD18-T Simple Vector,plasmids were extracted,identified by PCR and restriction enzyme digestion.Will cSZ1 gene and PMV361 vector by the restriction enzyme PvuⅡand ClaI double digestion,next DNA connection.Results by PT-PCR amplified cSZ1 gene size 940bp,connected to PMV361 vector,the successful construction PMV361-cSZ1.Conclusion The amplified Eimeria acervulina cSZ1 gene open reading frame and constructed pMV361-cSZ1 express vector,to research a chicken coccidiosis vaccine to provide an initial experimental basis.