title: Heterologous protein expression in Trichoderma reesei using the cbhII promoter.
abstract: To express homologous or heterologous proteins in fungi, a protein expression system using the promoter of cellobiohydrolase II gene (cbhII) was constructed by generating an expression vector called pWEIIF00. The obtained vector possesses the left and right borders, a hygromycin phosphotransferase B selective marker and a strong promoter and terminator of cbhII from Trichoderma reesei. It can easily undergo random recombination. The applicability of the vector was tested by red fluorescent protein gene (DsRed2) expression detection in T. reesei Rut C30. Using this system, a recombinant Cel5A variant, N342R (Qin et al., 2008), was then selected to express in Rut-C30. Compared to that of the parent strain, integration of the N342R gene resulted in 31.09% increased carboxymethyl-cellulose-degrading (CMCase) activity at pH 5.0 and 56.06% increased activity at pH 6.0. The increased CMCase activity of the recombinant strains would be beneficial for its application uses in multiple industries. The vector constructed in this study can used in fungi to produce industrial proteins.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Fanju
authors.initials: F
authors.lastname: Meng
authors.forename: Dongzhi
authors.initials: D
authors.lastname: Wei
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
mesh_terms: D002480:Cellulase / Q000378:metabolism; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D005456:Fluorescent Dyes / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014242:Trichoderma / Q000235:genetics*
chemical_list: D017931:DNA Primers; D005456:Fluorescent Dyes; D011994:Recombinant Proteins; C056373:endoglucanase 2; D002480:Cellulase; D043366:Cellulose 1,4-beta-Cellobiosidase
keywords: Cellulase; Endoglucanase; Trichoderma reesei; cbhII Promoter