Investigation of Trichoderma harzianum XynII gene expression in Escherichia coli and tobacco

• Main Authors: Ming-chan Ciou, 邱名禪
• Other Authors: none
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/47832765783153780150

碩士 === 國立臺南大學 === 生物科技學系碩士班 === 99 === Plant cell walls are mainly composed of cellulose, hemicellulose and lignin. Xylose is the main building block for hemicellulose . Xylose,connected by β-1,4 bonds to form the polymer. Xylanase to improve the quality of the paper in the paper industry. In addition to use in food processing and animal husbandry, with high economic value. In this study, xylanases extracted from the Trichoderma harzianum EST323. Xylanase genes of cloning from the cDNA library. Xylanase to produce in E. coli and tobacco, to further explore its biological activity and yield. In the E. coli , the pET26 expression vector, respectively to do bonding by xyn II, xyn II: Rev4S, xyn II: Rev4 , and transformation to E. coli strain BL21. The E. coli by IPTG induced to mass production xylanase. In the tobacco, by the Agrobacterium strain EHA105 transformation of tobacco and transgenic tobacco by tissue culture to mass production. The Results: Restriction enzyme analysis showed xyn II gene has been ligation in the pET26 expression vector. Congo red staining showed that the strains with xylanase activity. Histochemical analysis and PCR analysis showed that exogenous xyn II gene into transgenic tobacco genome. SDS-PAGE analysis showed molecular weight of about 22 kDa and 24 kDa. Western blot to the recombinant protein was successfully induced xyn II protein. How to improve yield xyn II protein in E.coli and in tobacco is the subject of future study.