文献类型： 会议论文
第一作者： Rihe Peng
作者： Rihe Peng 1 ; Aisheng Xiong; Xian Li; Huiqin Fan; Quanhong Yao;
作者机构： 1.Shanghai Key Laboratory of Agricultural Genetics and Breeding, Agro-Biotechnology Research Center, Shanghai Academy of Agricultural Sciences, Shanghai, China 201106
关键词： Synthetic crylA (c);successive PCR;expression;promoter, Escherichia coli
会议名称： Promoting global innovation of agricultural science & technology and sustainable agriculture development,Session 4
主办单位：
页码： 277-281
摘要： Successive polymerase chain reaction (PCR) was used to modify and synthesize a DNA fragment encoding the activated toxin domain of Cry IA (C) produced by Bacillus thringensis for the preferential expression in Pseudomonas fluorescence. The nucleotidesequence of the synthesized gene had 66.8% homology with the wide type, 1.8 Kb crylA (c) gene. The differences in 614 bases resulted in a total of 496 codon changes to be preferential to Pseudomonas spp. A TAG stop codon was introduced downstream of the1.8 kb nucleotide fragment. All secondary hairpin structure and potential unstable element were eliminated, and the G+C content was increased from 37.3% to 64%. The synthetic gene was introduced into vector pYP2Q01 for expression in Escherichia coll DH5tx under the control of tac promoter. At optimal conditions, the coding 66Kda protein was amounted to 20% of the total cellular protein.
分类号： S
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