Cloning and Prokaryotic Expression Character Analysis of AhRab7 Genes from Peanut (Arachis hypogaea L .)
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    Abstract:

    The full length cDNA of AhRab7-1、AhRab7-2 was isolated from HuaYu 20 (Arachis hypogaea L.) by RT-PCR and transferred into Escherichina coli to analyzed if the expression of the small G protein AhRab7 in E. coli was associated with the tolerance to high salt environment for E. coli. AhRab7-1 and AhRab7-2 were 872 bp and 816 bp, containing a 621bp and 618bp open reading frame and encoding 206 and 205 amino acids respectively. AhRab7-1 and AhRab7-2 were cloned into the expression vector pET-28a(+) with full-length ORF, then transferred into E. coli. After inducing by IPTG, the E. coli were treated with high salinity stresses and the function of the protein AhRab 7 could be tested. The tolerance activity assay showed that pET-28a-AhRab7-1 and pET-28a-AhRab7-2 had normal restructure enzyme activities and significantly released the salt tolerance of E. coli in LB medium with high NaCl concentration (5.5% ~10%). The control which had been transferred only the vector pET-28 did not showed the similar enzyme activity. The result suggested the expression of AhRab7 in E. coli could extremely improve the tolerance of salt for E. coli. The product proteins of AhRab7-1 and AhRab7-2 were detected by SDS-PAGE and the proteins were 23kDa as expected. Our researches could be used as a starting point for generation of plants tolerant to Saline-alkali and other abiotic stress in the future.

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History
  • Received:September 14,2012
  • Revised:December 27,2012
  • Adopted:March 11,2013
  • Online: June 07,2013
  • Published:
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