Molecular cloning, expression and purification of hyper-variable region in the hexon protein of human adenovirus

Abstract

Objective To clone and express the hyper-variable region in the hexon protein of human adenovirus, understand its structure and function, and lay a foundation for a genetic engineering vaccine candidate of human adenovirus. Methods We selected the target gene which was the hypervariable region of hexon, according to the model of human adenovirus hexon predicted by bioinformatics software.The corresponding primers were designed.The hexon, preserved by our team, was as a template to amplify the desired gene by PCR.The recombinant plasmid was constructed by directly cloning into the vector pQE32.The positive engineering vector, confirmed by the cleavage of endonuclease and nucleic acid sequence, was transformed into E. coli.M15 to express the His-tagged protein.Then the target protein was purified by Ni-Resin column affinity chromatography. Results The recombinant plasmid containing the hyper-variable region fragment in the hexon protein of human adenovirus was constructed.The conditions for the target gene expressing protein efficiently in E. coli M15 were found.We purified the target protein with a molecular weight of 188 000, which was consistent with the prediction by bioinformatics. Conclusion The fragment of hexon protein hyper-variable region is cloned, expressed and purified successfully. Key words: Human adenovirus; Hexon hyper-variable region fragment; Protein expression; Protein purification