Expression of Rs-AFP2 in Escherichia coli and Its Antifungal Assay
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    Abstract:

    Rs-AFP2 is an antifungal peptide from radish(Raphanus sativus).In this study,the full cDNA sequence of Rs-AFP2 gene was synthesized,and was sub-cloned in-frame to the 3′-terminus of the ORF of Glutathione S-transferase(GST)in the prokaryotic expression vector pGEX-4T-1.This allowed the recombinant protein GST-Rs-AFP2 to be highly expressed in cells of Escherichia coli.The recombinant protein GST-Rs-AFP2 could be expressed in the form of inclusion bodies after induction by isopropyl-β-D-thiogalactoside.Then,the GST-Rs-AFP2 protein was solubilized,denatured,refolded,and purified.The results of antifungal assay in vitro showed that the Rs-AFP2 protein exerted obvious inhibition against the growth of phytopathogenic fungi,such as Rhizoctonia solani,R.cerealis,Bipolaris sorokiniana,VerticilliuIll dahliae, and Fusarium graminearum.Therefore,Rs-AFP2 gene may be useful for improving the crop resistance against the above pathogenic fungi.

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