The Construction Of Filamentous Fungi Cellulase Gene Transformation System Mediated By Agrobacterium Tumefaciens And It's Application In The Character Study Of Cellulase

As the fuel was used more and more the traditional energy are at the danger of being used up.In order to make the ecomomic of the state grow healthly every state all of the whorld begain to look for the new substitutable renewable energy. Cellulose is the most ample renewable energy on earth.It take 40 percent of all the biomass on earth. So to use the cellulose sufficiently will be a good way to solvle the energy predicament.There are many methods to hydrolyze cellulose.Commonly aid and high temperature are used which result in unwilling byproduct and polluting the environment and damaging glucose.Now more attention has been taken to the enzymatic hydrolysis.But low efficiency of cellulose astrict it's extensive application. Nowadays gene engineering technology and protein engineering technology are practiced to cellulose modify and epression.A high efficiency filamentous fungil express system is needed to express cellulase. E.coli and P. pastoris had been proved to be unfit expression systems.This paper we practise Agrobacterium tumefaciens transformation system for transformation. Based on this system we transform the CBH I gene into the genome of the Neurospora crassa.And we practise to make deeper application of this system.Under the help of Dr. Chandrika Mulakala,we use the program Swiss-pdb viewer,PyMOL ect to analyse the structure of the CBD and the mechanism of the CBH I during cellulose biodegradation. After analysis we got several amino acid which may touch to the cellulose durying hydrolysis reaction. To prove the hypothesis we have mutated those sites by in vitro site-directed mutagenesis.Then using the established system we transform tow mutated gene to the Neurospora crassa.By the pilot study the result do not prove our hypothesis well. It needs more work and further analysis.