Cloning and expression of bovine pancreatic ribonuclease A gene and preliminary characterization of the recombinant enzyme

Zhang Quan; Yuan Yan; Zhu HongFei; Zhang XinYu; Sun HuaiChang

Scientia Agricultura Sinica 39(6): 1248-1252

2006


ISSN/ISBN: 0578-1752
Accession: 013051074

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Abstract
This study was conducted to determine the use of recombinant protein in the preparation of plasmid DNA for gene therapy and immunization. The isolated RNA from bovine pancreas and ribonuclease A gene were amplified. RT-PCR product was subcloned into pGEM-T vector for sequencing and subcloned into prokaryotic expression vector pEZZ18 for the expression of EcoRI/BamHI fragment. Results revealed that the predicted 28 kD fusion protein was expressed in the periplasmic space and the isolated RNA and plasmid were intact. It is concluded that the recombinant protein can hydrolyze the RNA and be used in purifying plasmid DNA.