核酶M1GS-T6对HCMV UL97基因RNA片段体外切割作用

Targeted in vitro cleavage of HCMV UL97 mRNA segments by M1GS-T6 ribozyme

目的:研究M1GS核酶对HCMV UL97 mRNA的体外切割作用。方法:针对HCMV UL97 mRNA T6位点设计与之互补的引导序列(Guide Sequence,GS),将其共价结合至大肠杆菌核酶P催化亚基(M1 RNA)的3′末端,构建M1GS-T6核酶,并用其对UL97基因亚克隆片段转录产物进行体外靶向切割实验。结果:核酶M1GS-T6具备特异性切割靶分子UL97mRNA的能力。结论:核酶M1GS-T6具备特异性切割活性,为进一步研究HCMV病毒基因功能和治疗提供了新的途径。

Aim： To study the cleaving activity of M1GS ribozyme targeting HCMV UL97 mRNA in vitro. Methods： The M1GS-T6 ribozyme was constructed by linking the catalytic RNA subunit of RNase P from Escherichia coli （M1 RNA） covalently to a guide sequence （GS） that is complementary to the mRNA coding sequence of UL97, which encode a phosphotransferase of HCMV. Results： The UL97 mR- NA was specifically cleaved by M1GS-T6 in our intro system. Conclusion： The M1GS-T6 we constructed showed specific enzyme activity on cleaving UL97 mRNA. It suggests a potential application as a novel antiviral agent.