Expression,purification and identification of truncated fusion protein (F) from Lanzhou strain of human respiratory syncytial virus

Objective The study aims to clone and express the fragment of the gene encoding fusion protein(F) from Lanzhou strain of human respiratory syncytial virus(HRSV) and to purify and identify the expressed product.Methods PCR technique was employed to amplify the gene fragment of F protein from Lanzhou strain of RSV.The amplicon was ligated with pET-42b(+) and the plasmid was transformed into E.coli Rosetta for expression.The transformants were induced by IPTG to express the recombinant protein,which was subjected to purification via ProbBondTM Ni-chelating resin column.The expressed protein was analyzed by SDS-PAGE and its reactogenicity was identified by Western-blot analysis.Results The DNA fragment with length of 951 bp was amplified.Both restriction enzyme analysis and sequencing revealed that the recombinant plasmid was correctly constructed.The molecular weight of the expressed protein was 68 710,its content was as high as 7%.After purification,the purity of the recombinant protein reached 80%.Conclusion The expressing vector expressing F protein from Lanzhou strain of RSV was constructed successfully.The recombinant protein was expressed and purified and used for diagnosis of sera and development of kit used for detection of HRSV infection.