关键词: 绿色木霉, TvALP-linker-TvRBL融合基因, 信号肽, 原核表达

Abstract: Fusion gene were constructed by linking the TvALP and TvRBL genes with a flexible chain（linker）. Bioinformatics analysis showed that the fusion gene contained a signal peptide. The TvRBL、TvALP and ?TvALP genes were cloned by RT-PCR from the total RNA of Trichoderma viride mycelium, which in turn were cloned into expression plasmid pET30a to construct prokaryotic expression plasmids pET30a-TvALP-linker-TvRBL and pET30a-?TvALP-linker-TvRBL, then these two expression plasmids were transformed into E.coli BL21（DE3）pLysS, and protein expression were induced by IPTG. SDS-PAGE was used to analyze the expression of the fusion protein. The result showed that expression plasmid pET30a-?TvALP-linker-TvRBL was obtained expression in E.coli BL21（DE3）pLysS, which was a specific band at about 60 kD in size and identical with the expected molecular weight of the fusion protein.

Key words: Trichoderma viride, TvALP-linker-TvRBL fusion gene, signal peptide, prokaryotic expression