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

LIN Lin; WU Yun—feng”; CUI Xiao-feng

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February 2003

LIN Lin, WU Yun—feng” and CUI Xiao-feng. Cloning and Prokaryotic Expression of groEL Gene of Endosybiotic Bacterium from rhopalosiphum maize[J]. Virologica Sinica, 2003, 18(1): 54-57.

Citation: LIN Lin, WU Yun—feng”, CUI Xiao-feng. Cloning and Prokaryotic Expression of groEL Gene of Endosybiotic Bacterium from rhopalosiphum maize .VIROLOGICA SINICA, 2003, 18(1) : 54-57.

玉米蚜体内参与传毒的共生菌groEL基因的克隆和原核表达

1.
西北农林科技大学植物保护学院，陕西杨陵712100

摘要

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墓因全长为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一30a表达出69KDa的融合蛋白，二者均有较高的表达量。
- 玉米蚜：内共生菌
- , GroEL
- , 传毒：表达载体

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

1.
Department of Hant protection，North-West Sci-Tech University of Agricultare and Forestry,Yangling 712100， China

Abstract

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 sequeneee levels． this protein was identified as GroEL protein of endosymbiotic bacterium (Buchnera sp．) of aphid．The results of nucleotides sequencing indicated that the full length of groEL genes f GeneBank accessio number iS AF3 87863) iS 1 647bp and code 548 amino acids．Th e cloned groEL genes of and Rhopalosiphum maize were ligated into pBV22 1，pET30a and pTrcHisA vector for expressing The results suggested that 63 KDa aim proteins can be espressed using pBV22 1．When pET一30a used，the expressed fusion proteins have a molecular weight of 69 KDa，and both have a higher expression quantiy
- Rhopalosiphum maize
- , Endosymbiotic bacterium
- , GroEL

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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

1. Department of Hant protection，North-West Sci-Tech University of Agricultare and Forestry,Yangling 712100， China

Abstract: 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 sequeneee levels． this protein was identified as GroEL protein of endosymbiotic bacterium (Buchnera sp．) of aphid．The results of nucleotides sequencing indicated that the full length of groEL genes f GeneBank accessio number iS AF3 87863) iS 1 647bp and code 548 amino acids．Th e cloned groEL genes of and Rhopalosiphum maize were ligated into pBV22 1，pET30a and pTrcHisA vector for expressing The results suggested that 63 KDa aim proteins can be espressed using pBV22 1．When pET一30a used，the expressed fusion proteins have a molecular weight of 69 KDa，and both have a higher expression quantiy