[Cloning of gene related to salt tolerance from Sinorhizobium fredii RT19 and its expression in Escherichia coli].
A 4.4 kb DNA fragment related to salt tolerance containing three open reading frames was isolated from the gene library of S. fredii strain RT19. By subcloning and functional analysis, only ORF2 related to salt tolerance was obtained. The ORF2 was ligated to expression vectors pThioHisA, B and C, respectively, and recombinant expression vectors pGA, pGB and pGC containing 1.5 kb DNA fragment related to salt tolerance were constructed. These recombinant expression vectors were transformed into E. coli DH5 alpha. Inducing by IPTG and analyzing with SDS-PAGE, it was found that the fusion protein encoded by pGC was expressed, and its molecular weight was equal to the sum of thioredoxin encoded by trxA and ORF2 putative protein molecular weight. The Western blot demonstrated that the target gene was successfully expressed in E. coli.
Ge
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SC
Department of Microbiology, College of Biological Sciences, China Agricultural University, Beijing 100094, China.
Liu
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YN
Yang
S S
SS
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D020579:Sinorhizobium / Q000235:genetics*; D012965:Sodium Chloride / Q000494:pharmacology*
D012965:Sodium Chloride