[Construction and identification of the recombinant prokaryotic expression plasmid containingbetapep-25 peptide].

ABSTRACT

AIM: To synthesize antiangiogenic peptide fragment of betapep-25, to construct and identify the recombinant prokaryotic expression plasmid containing betapep-25 peptide. METHODS: The fragment encoding betapep-25 peptide was designed and synthesized artificially and was cloned into vector pGEM-T easy after being identified by sequencing. After being digested by Nae I and BamH I, T-betapep-25 peptide fragment was cloned into recombinant vector pBV220-NT4, which was digested by Nae I and BamH I. The constructed recombinant prokaryotic expression plasmid pBV220-NT4-betapep-25 was digested using BamH I, EcoR I and BamH I, respectively. RESULTS: The sequcence of betapep-25 synthesized artificially and analyzed by digestion was consistent with the published results. Fragments of 4,030 bp, 364 bp and 3,666 bp were obtained after digestion of recombinant prokaryotic expression plasmid pBV220-NT4-betapep-25 using BamH I, EcoR I and BamH I, respectively, which demonstrated the successful cloning and construction of recombinant prokaryotic expression plasmid. CONCLUSION: The successful construction recombinant prokaryotic expression plasmid expressing betapep-25 provides a foundation for further research on its mechanism of anti-tumor activity in vivo and in vitro.