Cloning of Endoglucanase Gene from Trichoderma harzianum in E . coli

Recycling of agro-industrial wastes to produce high quality protein is an efficient and economical way to solve the problem of deficiency in good quality protein. Cellulases are attractive source for utilization of the agro-industrial waste materials. Endoglucanase (EC 3.2.1.4) was isolated from Trichoderma harzianum (E-58 Strain). It was grown on Vogel's medium with different carbon sources. Maximal production of endoglucanase gene was achieved at 28°C, pH 5.5 under continuous shaking at 120 rpm for 5 days. Glucose repressed the synthesis of enzyme whereas carboxymethylcellulose produced the enzyme in substantial amounts. RNA was isolated from mycelia of T. harzianum. First strand of cDNA was synthesized using oligo dT (18) primer and subjected to PCR with sequence specific primers. The amplified products were purified through agarose gel electrophoresis and ligated into pUC18. The plasmids containing endoglucanase genes were transformed into E. coli for further characterization.

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