Molecular Cloning and Characterization of a Putative cDNA Encoding Endoglucanase IV from <Emphasis Type="Italic">Trichoderma Viride</Emphasis> and its Expression in <Emphasis Type="Italic">Bombyx Mori</Emphasis> |
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Authors: | Xing-hua Li Peng Zhang Shuang Liang Fang Zhou Mei-xian Wang Roy Bhaskar Firdose Ahmad Malik Yan-shan Niu Yun-gen Miao |
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Institution: | (1) Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, 310029, People’s Republic of China;(2) College of Animal Sciences, Zhejiang University, Hangzhou, 310029, People’s Republic of China; |
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Abstract: | The development of cellulase production technology has greatly contributed to the successful use of cellulosic materials as
renewable carbon sources. In this study, a putative endoglucanase IV (EG IV) complementary DNA was cloned from the mycelium
of a strain of the filamentous fungus Trichoderma viride using a PCR-based exon-splicing method and expressed in both a silkworm BmN cell line and in silkworm larvae. Western blot
analysis detected a band of 42 kDa in BmN cells after infection with a recombinant mBacmid/BmNPV/EG IV baculovirus. Sequence
alignment analysis of the T. viride EG IV gene showed two domains that were highly conserved with glycosyl hydrolases and a funga-type cellulose-binding domain.
Analysis of variance showed that silkworms infected with recombinant baculoviruses exhibited significantly higher enzyme activity
that was 48.84% higher than silkworms infected with blank baculoviruses and 46.61% higher than normal silkworms. The expressed
bioactive EG IV was also stable at the pH range from 5.0 to 10.0. The availability of large quantities of bioactive EG IV
in silkworm provided a possibility to produce cellulase transgenic silkworm, which express bioactive cellulase specially in
its digestive tract and improve its metabolism efficiency of mulberry leaves. Its application in the sericulture industry
may be very promising. |
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