title: Molecular cloning and characterization of a putative cDNA encoding endoglucanase IV from Trichoderma viride and its expression in Bombyx mori.
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.
authors: Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou 310029, People's Republic of China.
authors: Xing-Hua
authors: XH
authors: Li
authors: Peng
authors: P
authors: Zhang
authors: Shuang
authors: S
authors: Liang
authors: Fang
authors: F
authors: Zhou
authors: Mei-Xian
authors: MX
authors: Wang
authors: Roy
authors: R
authors: Bhaskar
authors: Firdose Ahmad
authors: FA
authors: Malik
authors: Yan-Shan
authors: YS
authors: Niu
authors: Yun-Gen
authors: YG
authors: Miao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D012831:Bombyx / Q000235:genetics* / Q000821:virology; D002460:Cell Line; D002480:Cellulase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics; D005091:Exons / Q000235:genetics; D015870:Gene Expression; D007814:Larva / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D012326:RNA Splicing; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D004274:DNA, Recombinant; D002480:Cellulase