Pesquisa Agropecuária Brasileira
Print version ISSN 0100-204X
LOMBARDI, Ricardo; HARAKAVA, Ricardo and COLARICCIO, Addolorata. Cloning and purification of Banana streak OL virus coat protein fragment. Pesq. agropec. bras. [online]. 2010, vol.45, n.8, pp.811-817. ISSN 0100-204X. http://dx.doi.org/10.1590/S0100-204X2010000800006.
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.
Keywords : Badnavirus; Escherichia coli; Musa; antisera; banana plant; recombinant protein expression.