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Cellulase Gene Clone, Genetic Transformation and the Research on Cre/loxP System

Author: GeXiGeTu
Tutor: CiZhongLing
School: Inner Mongolia Agricultural University
Course: Tree Genetics and Breeding
Keywords: endo-glucanase gene (EGⅡ) exo-glucanase gene (CBHⅡ) emale sterility endoplasmic reticulum retention signal tapetal-specific promoter Cre/loxp recombination system
CLC: Q78
Type: Master's thesis
Year: 2009
Downloads: 109
Quote: 1
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Abstract


A large number of cellulose become essential in the lives of human being with the use of cellulose enzymes. It has great importance to solving the world’s energy crisis, food shortages and environmental pollution, etc. In addition, bio-security issues have gradually become a topic of global concern with the development of modern biotechnology, especially recombinant DNA technology and the genetic engineering. Deleting the marker gene (herbicide-resistant gene) using Cre/loxP system is vital to bio-security issues. In this paper, the specific work as follows:1. Using RT-PCR method, endo-glucanase gene (EGⅡ) and exo-glucanase gene (CBHⅡ) have been cloned from the green trichoderma. Exo glucanase gene (CBHⅡ) has three introns and four exons, while the endoglucanase gene (EGⅡ) no intron.2. Through adding endoplasmic reticulum stranded signal in EGⅡgene, a P3300-35S-EGⅡ-Tnos vector is built and then transferred to tobacco. 7 transgenic plants have been identified by PCR.3. Using the Cre/loxP system and the male sterility gene to build expression vectors, delete the marker gene (herbicide-resistant gene) and transfer plant expression vector P3300 Cre/lox-Osg6B-Barnase-Tnos into bent grass. 6 transgenic plants pass though PCR identification.

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CLC: > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering)
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