阴道毛滴虫琥珀酰辅酶A合成酶基因的克隆与序列分析

Cloning and sequence analysis of α-succinyl CoA synthetase B gene from Trichomonas vaginalis

目的获取阴道毛滴虫琥珀酰辅酶A合成酶(α-SCSB)全基因,并对其进行序列分析。方法利用PCR技术扩增阴道毛滴虫琥珀酰辅酶A合成酶(α-SCSB)全基因,并将其插入pMD18-T载体,酶切鉴定后进行序列测定及分析。结果PCR扩增获得α-SCSB全长基因,大小为1381bp,与国外发表的阴道毛滴虫琥珀酰辅酶A合成酶(α-SCSB)的基因同源性为100%。结论本研究获得了阴道毛滴虫琥珀酰辅酶A合成酶(α-SCSB)启动子,为下一步研究α-SCSB启动子的功能及构建阴道毛滴虫DNA稳定转染载体奠定了基础。

Objective To clone and analyze the total α-succinyl CoA synthetase B （α-SCSB） gene from Trichomonas vaginalis. Methods The gene was amplified by PCR and inserted into pMD18 T vector. The recombinant plasmid pMD/α-SCSB was identified by restriction enzyme digestion and sequenced. Results The sequence of α-SCSB was 1 381 bp. It was proved that the sequence of α-SCSB gene was just the same as that of the T. vaginalis α-SCSB gene reported abroad. Conclusion It is evident that the α-SCSB gene promoter was successfully obtained in the present study. This is a basis to study the function of α-SCSB gene promoter and construct DNA stable transfection vector of T. vaginalis.