变异链球菌LuxS基因表达载体的构建

Construction of vector expressing LuxS gene

目的构建可用于变异链球菌表达系统的含LuxS基因的原核表达载体。方法应用聚合酶链反应(PCR)方法分别从变异链球菌的DNA、载体pEGFP-N1中扩增出LuxS基因的启动子(pLuxS)和绿色荧光蛋白(gfp)基因片段,经回收、纯化后,利用分子克隆技术分别克隆入载体pUC19中。结果通过对重组质粒pUC19-pLuxS-gfp的酶切图谱和DNA序列测定分析证实插入片段序列正确。结论成功构建出原核表达重组质粒pUC 19-pLuxS-gfp为下一步的功能研究奠定了基础。

Objective To obtain a prokaryotic expression vector containing LuxS gene of Streptococcus mutans. Methods LuxS promoter and gfp region were amplified by PCR from S. mutans DNA and pEGFP-N1 vector, respectively. Then they were cloned into the pUC19 vector following routine procedures by using cloning technique. Results Identification by enzyme digestion, PCR and sequencing confirmed the valid- ity of the recombinant vector pUC19-pLuxS-gfp. Conclusion The successful construction of the recombinant plasmid pUC 19-pLuxS-gfp will be further studied in this field.