Expressing Cellulase Gene In Transgenic Rice Seed

Besides to the grain, the biomass resources mainly refers to lignocelluloses class farming and forestry reject. There are extremely abundant biomass resources. In addition, the biomass resources research and development is becoming one of the world significant popular topics. The principle constituent of the biomass is the cellulose, the hemicellulose and the lignin, in which, the cellulose and the hemi-celluloses account for 70% above among the biomass total quantity. Many types of cellulase not only hydrolyse cellulose, but also exhibit a high activity in depolymerisation of 1,3-1,4-β-glucans by hydrolysing the 1,4-β-glucosidic linkage. The application of cellulase is extremely widespread. To produce the cellulase using the plant as the bioreactor has the merit of low price, bear storing, not suitably pollution and so on. It has been previously reported that the cellulase could be highly expressed in barley endosperm.This experiment though Agrobacterim-mediated Transformation in order to transform the cellulase gene EGI into rice, which is from Trichoderma reesi.. Driven by the storage protein promoter , it could express in rice endosperm. First of all, the plant expression the cellulase is obtained. Afterward, it has been carried on the PCR appraisal to the plant and takes the active test on expressed the cellulase. It disposes the substrate with each kind of different pH value cushion fluid. It takes the active test in different temperature and pH value, and then it could obtain the temperature and pH value grad curve. It also ascertains the most suitable enzyme condition to the cellulose and hemi-celluloses substrate. The most suitable condition of the enzyme to the cellulose substrate is 45℃, pH4.6; meanwhile, the most suitable condition of the enzyme to the hemicellulose substrate is 45℃, pH6.0. Lives under the most suitable condition to the cellulose substrate enzyme is the 4.9U/g seed, hemicellulose substrate enzyme lives under the most suitable condition is the 6.2U/g seed. Moreover, it could examine the cellulose enzymatic activity, indicating that gene in T1 paddy rice seed has transmissibility.