Xylan is the major component of hemicellulose and should be fully utilized to improve the efficiencies of a biobased economy. Xylanases have received great attention in the development of environment friendly technologies in the paper and pulp industry. The use of xylanases could greatly improve the overall lignocellulosic materials for the generation of liquid fuels and chemicals. Trichoderma harzianum E-58 was grown in Vogel`s medium with different carbon sources such as glucose, maltose, carboxymethylcellulose (CMC), wheat bran and xylan at 28°C with constant shaking at 150 rpm to check the induction of xylanase. Expression of xylanase was induced maximally by xylan and repressed by glucose. The pH and temperature optima of the crude xylanase were 5 and 60°C, respectively. The apparent Km and Vmax values of the crude xylanase using Birchwood xylan as a substrate were 2 mg m/ L-1 and 3.35 µmol min-1 mg-1, respectively. RNA wass isolated from mycelia of T. harzianum by using Trizol reagent. cDNA was synthesized by using cDNA synthesis kit. The subsequent PCR products were purified through agarose gel electrophoresis and ligated into pUC18. The recombinant vectors containing the xylanase gene were transformed into E. coli DH I OB for characterization.