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Enhanced expression of a recombinant malaria candidate vaccine in Escherichia coli by codon optimization

机译:通过密码子优化增强重组疟疾候选疫苗在大肠杆菌中的表达

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摘要

This study was conducted to compare the expression of three constructs of a multistage candidate vaccine (FALVAC-1) against Plasmodium falciparum in an Escherichia coli system: a synthetic gene with P. falciparum codons. a synthetic gene with optimized E coli codons, and a synthetic gene with P. falciparum codons co-transformed with a RIG plasmid, which encodes three tRNAs (AG(A/G). ATA. GGA) that recognize rare E. coli codons. The expression of the protein increased at least threefold with codon optimization. The presence of the RIG plasmid in the co-transforming cells did not significantly increase the expression level of the gene with P. falciparum codons. The growth of cells transformed by the construct with P. falciparum codons was significantly slower than that of cells transformed by the construct with optimized E coli codons after induction of protein expression with IPTG. The cells containing the non-codon optimized gene co-expressed with RIG plasmid had the slowest growth at all time points in culture. Thus, codon optimization significantly increases the yield of P. falciparum candidate vaccines in the Each expression system. (C) 2003 Elsevier Inc. All rights reserved. [References: 44]
机译:进行这项研究的目的是比较大肠杆菌系统中针对恶性疟原虫的多阶段候选疫苗(FALVAC-1)的三种构建体的表达:一种具有恶性疟原虫密码子的合成基因。一个带有优化大肠杆菌密码子的合成基因,以及一个带有恶性疟原虫密码子的合成基因,该基因与一个RIG质粒共转化,该质粒编码三个识别稀有大肠杆菌密码子的tRNA(AG(A / G)。ATA.GGA)。蛋白质的表达随着密码子优化而增加至少三倍。在共转化细胞中,RIG质粒的存在并没有显着提高恶性疟原虫密码子的基因表达水平。用IPTG诱导蛋白质表达后,由恶性疟原虫密码子构建体转化的细胞的生长明显慢于由具有优化大肠杆菌密码子的构建体转化的细胞的生长。在培养的所有时间点,含有与RIG质粒共表达的非密码子优化基因的细胞生长最慢。因此,密码子优化显着提高了每个表达系统中恶性疟原虫候选疫苗的产量。 (C)2003 Elsevier Inc.保留所有权利。 [参考:44]

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