Preparation of agrobacterium tumefaciens strains containing PEX2B plasmids carrying gene coding for polysaccharide monooxygenases

Abstract

To minimize environmental pollution and ensure energy security when fossil fuel sources are oil depleted, most countries in the world have been turning to use biofuels. Polysaccharide monooxygenases (PMOs) which degrade tenacious polysaccharides including cellulose in the plant is paid special attention to by scientists as a solution for the biofuel. In this study, primers designing for AA9 genes amplification and pEX2B plasmid linearization following Gibson Assembly method which0 turns to obtain pEX2B plasmid containing AA9 genes that can be used for Aspergillus oryzae transformation. After the period of the thesis, not only primers to amplified AA9 genes was designed, but also the cloning AA9 genes into pEX2B vector protocol was created. Moreover, pEX2B-AA9 vector transformation was performed successfully. This study finds a way to improve the ability to apply PMOs to the industry. This topic has shown the potential for using A. tumefaciens to improve the ability of PMOs application to be used into the energy industry and as a premise for future PMOs expression through A. oryzae.