The Construction, Expression and Purification of Therapeutic Adjuvant-free protein Vaccine—HPV16Z-Hsp65-E6/E7

China Biotechnology

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The Construction, Expression and Purification of Therapeutic Adjuvant-free protein Vaccine—HPV16Z-Hsp65-E6/E7

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Abstract Objective: To develop a therapeutic adjuvant-free protein vaccine against HPV16 which is closely related to cervical cancer of China. Method: We got first the E6/E7 gene by PCR technology from HPV16z virus strain isolated in the high-risk cervical cancer area of Shanxi Province of China in1990s, and again got the gene segment of Hsp65 from BCG by the same method, we mutated the transforming codes in sequences of HPV 16 E6/E7 genes and thus constructed the expression vector pET28a-Hsp65-E6/E7, expressed the Hsp65-E6/E7 fusion protein in E.coli BL21(DE3) strain and researched optimal protein purification procedures. Results: The expression vector pET28a-Hsp65-E6/E7 was constructed successfully and E6/E7 gene was mutated correctly. Hsp65-E6/E7 fusion protein was renatured and purified on the affinity chromatography column simultaneously. The protein purity achieved 95% after the anionic exchange chromatograph purification. conclusions: This research laid a foundation for further functional study of the therapeutic adjuvant-free protein vaccine -Hsp65-E6/E7.

Key words： cervical cancer HPV16 therapeutic vaccine

Received: 13 June 2006 Published: 25 December 2006

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Cite this article:

. The Construction, Expression and Purification of Therapeutic Adjuvant-free protein Vaccine—HPV16Z-Hsp65-E6/E7. China Biotechnology, 2006, 26(12): 40-44.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I12/40

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