Biotechnology Bulletin ›› 2015, Vol. 31 ›› Issue (1): 131-137.doi: 10.13560/j.cnki.biotech.bull.1985.2015.01.020

Previous Articles     Next Articles

Construction and Prokaryotic Expression of Trichoderma viride TvALP-linker-TvRBL Fusion Gene

Xu Yangyu1,2, Wen Shijie, Li Haifen, Li Ling2, Liang Xuanqiang   

  1. (1. Crops Research Institute,Guangdong Academy of Agricultural Sciences,Guangzhou 510640;2. College of Life Science,South China Normal University,Guangzhou 510631)
  • Received:2014-05-22 Online:2015-01-09 Published:2015-01-10

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