Construction of the Bivalent-expression Vector for Trichoderma spp.

doi:10.11924/j.issn.1000-6850.2013-1905

Abstract

Abstract: Bivalent-expression vector of chitianse gene andβ-1,4-glucanase gene lays the foundation for the transformation of Trichoderma spp.. The gene fragment was obtained by digesting of restriction endonuclease, and then lagased into expression cassette by T4 DNA ligase to form the bivalent-expression vector. The vector was verified by using PCR, digestion and nucleotide sequencing. The recombinant pC1302- C42 was constructed by cloned the chitinase gene chi42 into expression cassette with promoter CaMV35S and terminator PolyA, and the recombinant pC1300-2-G14 was constructed by cloning theβ-1,4-glucanase gene glu14 into expression cassette with promoter CaMV35S and terminator Nos, and then the bivalent-expression vector pC1300- 2- G14- C42 with the hygromycin B resistance gene was constructed through successful connection of 35S-chi42-PolyA and 35S-glu14-Nos. The bivalent-expression vector pC1300-2-G14-C42 can be directly applied to filamentous fungi genetic transformation, lays the foundation for screening broad spectrum engineering strain of Trichoderma spp..

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