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Molecular Cloning and Characterization of a Putative cDNA Encoding Endoglucanase IV from Trichoderma Viride and its Expression in Bombyx Mori

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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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Acknowledgments

The work was supported by the National Basic Research Program of China under grand number 2012CB114601 and the National Natural Science Foundation of China (number 30972141/C120110), the Key project of Zhejiang Government (number 2011C14006), and Chinese Universities Scientific Fund.

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Authors and Affiliations

  1. Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, 310029, People’s Republic of China

    Xing-hua Li, Peng Zhang, Shuang Liang, Fang Zhou, Mei-xian Wang, Roy Bhaskar, Firdose Ahmad Malik, Yan-shan Niu & Yun-gen Miao

  2. College of Animal Sciences, Zhejiang University, Hangzhou, 310029, People’s Republic of China

    Yun-gen Miao

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  1. Xing-hua Li
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  2. Peng Zhang
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  4. Fang Zhou
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  8. Yan-shan Niu
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Correspondence to Yun-gen Miao.

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Li, Xh., Zhang, P., Liang, S. et al. Molecular Cloning and Characterization of a Putative cDNA Encoding Endoglucanase IV from Trichoderma Viride and its Expression in Bombyx Mori . Appl Biochem Biotechnol 166, 309–320 (2012). https://doi.org/10.1007/s12010-011-9426-5

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  • DOI: https://doi.org/10.1007/s12010-011-9426-5

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