کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2824191 | 1161628 | 2012 | 5 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Construction of two vectors for gene expression in Trichoderma reesei Construction of two vectors for gene expression in Trichoderma reesei](/preview/png/2824191.png)
We report the construction of two filamentous fungi Trichoderma reesei expression vectors, pWEF31 and pWEF32. Both vectors possess the hygromycin phosphotransferase B gene expression cassette and the strong promoter and terminator of the cellobiohydrolase 1 gene (cbh1) from T. reesei. The two newly constructed vectors can be efficiently transformed into T. reesei with Agrobacterium-mediated transformation. The difference between pWEF31 and pWEF32 is that pWEF32 has two longer homologous arms. As a result, pWEF32 easily undergoes homologous recombination. On the other hand, pWEF31 undergoes random recombination. The applicability of both vectors was tested by first generating the expression vectors pWEF31-red and pWEF32-red and then detecting the expression of the DsRed2 gene in T. reesei Rut C30. Additionally, we measured the exo-1,4-β-glucanase activity of the recombinant cells. Our work provides an effective transformation system for homologous and heterologous gene expression and gene knockout in T. reesei. It also provides a method for recombination at a specific chromosomal location. Finally, both vectors will be useful for the large-scale gene expression industry.
► Two filamentous fungi Trichoderma reesei expression vectors pWEF31 and pWEF32 were newly constructed by us.
► Both vectors was confirmed by detecting the expression of DsRed2 gene in T. reesei Rut C30.
► The two vectors will be useful for large-scale gene expression industry.
Journal: Plasmid - Volume 67, Issue 1, January 2012, Pages 67–71