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| Expression of Codon-optimized Human Papillomavirus 16 L1 Gene in Two Insect Cells |
| GONG Ye-li1 WANG Lin2 GENG Jian-ling2 LIU Ying2 XIA Huan-zhang1 |
| (1 School of Life Science and Biopharmacology, Shenyang Pharmaceutical University, Shenyang110016, China)
(2 Shenyang Sunshine Pharmaceutical CO.LTD, Shenyang110027, China)
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Abstract Abstract Objective: To enhance the expression level of human papillomavirus (HPV)16 L1 through Baculovirus Expression System, in order to lay a foundation of development of prophylactic HPV vaccines. Methods:The codon usage of wild type HPV16L1 gene was optimized according to the codon bias of insect cells. In order to infect both Sf9 and High Five insect cells, the recombinant baculovirus was obtained using Baculovirus Expression System. The expressed product was identified by Western blot;The virus-like particles (VLPs) of HPV16 was confirmed by electron microscopy;And ELISA was used to evaluate the effect of the optimized gene as well as to determine the optimal expression condition. Results: The expressed protein, with a relative molecular mass of 56kDa, showed specific reaction with HPV16L1 antiserum;The intranuclear VLPs of HPV16L1 were observed by electron microscopy; The result of ELISA indicated that the expression level of optimized HPV16L1 gene was significantly higher than the wild type; And the optimal expression condition for High Five cells was determined as MOI 10 and expression phase 72 hours, in which the L1 protein was expressed at least 3 times higher than that in sf9 cells. Conclusion: Codon optimization could indeed enhance the expression level of HPV16L1 gene in insect cells.The significant advantage of High Five cells was worth attention.
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Received: 14 January 2009
Published: 28 July 2009
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