Cloning and purification of Banana streak OL virus coat protein fragment

Abstract

The objective of this work was to clone and to induce the expression of a fragment of Banana streak OL virus coat protein (BSOLV-CP) in Escherichia coli, as well as to purify the obtained recombinant protein. Two specific primers were used for the PCR‑amplification of approximately 390‑bp fragment of the codifying region of the BSOLV-CP central portion. The obtained fragment was cloned in pGEM-T Easy vector, subcloned in pQE-30 expression vector and transformed into competent E. coli M15 (pREP4) cells by heat shock. The protein expression was induced by isopropyl thiogalactopyranoside (IPTG) and the 14-kDa BSOLV-rcCP recombinant protein was detected in Western and Dot blotting. The expression of the BSOLV-rcCP protein enables new approaches to the obtention of antigens for the antisera production against BSOLV.