title: Construction of two vectors for gene expression in Trichoderma reesei.
abstract: 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.
authors: State Key Lab of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.
authors: Dandan
authors: D
authors: Lv
authors: Wei
authors: W
authors: Wang
authors: Dongzhi
authors: D
authors: Wei
mesh_terms: D060054:Agrobacterium / Q000235:genetics / Q000378:metabolism; D002480:Cellulase / Q000235:genetics / Q000378:metabolism; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000235:genetics; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D059765:Homologous Recombination; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic; D014242:Trichoderma / Q000235:genetics* / Q000378:metabolism*; D000097573:Red Fluorescent Protein
chemical_list: D002480:Cellulase; D043366:Cellulose 1,4-beta-Cellobiosidase; D004269:DNA, Bacterial; D008164:Luminescent Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); D000097573:Red Fluorescent Protein; C063242:hygromycin-B kinase