Cloning,Expression,Purification and Antimicrobial Activity of CrusSp from Scylla paramamosain

Hemolyph was collected from Scylla paramamosain. The total RNA extracted from the hemocyte sample was used to amplify the sequence encoding an open reading frame of a mature peptide of CrusSp by RT-PCR. The sequence was cloned into pMD18-T vector and subjected to DNA sequence analysis. The DNA of CrusSp was isolated from pMD18-T/CrusSp and was cloned into pET-32a(+) expression vector to allow expression of CrusSp as a fusion protein in E. coli OrigamiTM (DE3). The fusion protein could be purified effectively by His-Band resin chelating chromatography. The antimicrobial activities of CrusSp were studied by agar diffusion test. Analysis by 15% SDS-PAGE revealed that the molecular mass of CrusSp is 10.27 kDa. The antimicrobial activity revealed that CrusSp exhibited antifungal activity towards Trichoderma viride.