关键词: 日本血吸虫； 启动子； 基因操作； 报告基因； 克隆

Abstract:

Objective To evaluate the relevant promoter regions of four Schistosoma japonicum genes for expressing a lucifer? ase reporter. Methods The polymerase chain reaction（PCR）was used to amplify the promoter regions of four S. japonicum genes and then each PCR product was cloned into a pGlu?Basic vector according to the standard molecular procedures. These recombi? nant plasmids were either transfected into human HEK293 cells by using lipofectamine or introduced into cultured schistosomes by electroporation. Then，the luciferase activities were measured by using a dual luciferase reporter system in a luminometer. Re? sults Each promoter region of four S. japonicum genes was obtained and the corresponding recombinant vector containing the pro? moter region was successfully constructed. The transfection of the recombinant plasmids into the human HEK293 cells and cul? tured schistosomes resulted in a significant elevation of the luciferase reporter activity. Conclusions The promoter regions of four S. japonicum genes are obtained and the luciferase reporter genes driven by the four promoter regions are preliminarily evaluated. The study provides a foundation for the usage of these promoters for genetic manipulation in S. japonicum.

Key words: Schistosoma japonicum；Promoter；Genetic manipulation； Reporter gene； Clone