抗菌肽P113多拷贝表达载体的构建

Construction of Expression Vector of Antimicrobial Peptide P113

目的:构建唾液抗菌肽P113基因多拷贝串联重组体,并将其克隆到表达载体pET-28a。方法:根据大肠杆菌偏爱的密码子设计并合成人唾液抗菌肽P113基因,采用PCR技术构建P113基因的自融合多拷贝串联重组体polyP113,BamHⅠ和HindⅢ双酶切表达载体pET-28a和polyP113基因,将两者连接后转化大肠杆菌DH5α,氨苄青霉素筛选阳性菌落,PCR、酶切、测序鉴定重组质粒。结果:所构建的含有十拷贝P113基因的重组体正确克隆到表达载体pET-28a上,无突变。结论:成功构建含十拷贝唾液抗菌肽P113基因表达框的大肠杆菌表达载体。

Objective： To construct the tendem-repeated multiple copy recombinant for gene Salivary Antimicrobial Peptide P113 and then clone the gene to expression vector pET-28a.Methods： According to the optium codens of E.coli,we designed and synthesized the P113 gene PCR technique was used to construct the tendem-repeated multiple copy recombinant polP113,pET-28a and polyP113 were both digested by BamHⅠand HindⅢ,the digested pET-28a and polyP113 were linked and transformed to Ecoli DH5α,and the positive colonies were screened by ampicillin,and identified by PCR,digestion and sequencing.Result： The recombinant made up of ten copy P113 gene were correctly inserted into expression vector pET-28a without mutation.Conclusion： The expression vector of E.coli containing ten copies of Salivary Antimicrobial Peptide P113 was constructed successfully.