玉米蚜体内参与传毒的共生菌groEL基因的克隆和原核表达

Cloning and Prokaryotic Expression of groEL Gene of Endosybiotic Bacterium from Rhopalosiphum maize

以玉米蚜杨凌生物型为材料,设计特异性引物采用PCR的方法在国内首先克隆了一种玉米蚜体内参与传毒的共生菌groEL基因,序列测定结果表明:玉米蚜杨凌生物型共生菌groEL基因全长为1647bp.编码548个氨基酸,登录Genebank,序列号为AF387863。构建了该基因的原核表达载体,用pBV221表达出63KDa的非融合目的蛋白,用pET-30a表达出69KDa的融合蛋白,二者均有较高的表达量。

Cloned one virus binding protein from Rhopalosiphum maize Yangling biotype through PCR technique, using designed specific primers based on this protein partially sequence. In nucleotide and amino acid sequenece levels, this protein was identified as GroEL protem of endosymbiotic bacterium (Buchnera sp.) of aphid.The results of nucleotides sequencing indicated that the full length of groEL genes (GeneBank accessio number is AF387863) is 1647bp and code 548 amino acids. The cloned groEL genes of and Rhopalosiphum maize were ligated into pBV221, pET30a and pTrcHisA vector for expressing The results suggested that 63 KDa aim proteins can be espressed using pBV221. When pET-30a used, the expressed fusion proteins have a molecular weight of 69 KDa, and both have a higher expression quantiy.