[The expression and preliminary evaluation of HPV6bL2deltaN360E7E6 fusion protein in E. coli for genital warts]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2010 Jun;24(3):171-4.
[Article in Chinese]

Abstract

Objective: To express HPV6bL2deltaN360E7E6 fusion protein in E. coli and preliminarily evaluate its immune effect.

Methods: Three HPV6b gene fragments, which were L2(1-360 bp), E7 and E6, were fused by overlapping PCR, then were inserted into a prokaryotic expression vector and expressed in E. coli. C57BL/6 mice were immunized with purified fusion protein plus Al (OH)3 and/or CpG adjuvants through intramuscular route, the cellular and humoral immune responses were detected by IFN-gamma ELISPOT and ELISA respectively.

Results: Protein plus CpG adjuvant could induce the strongest cellular immune response to E7 and E6, high antibody titer against L2 could be detected in all immunized groups but there were no significant difference among these groups.

Conclusions: HPV6bL2deltaN360E7E6 gene was successfully cloned into pQE30 vector and expressed in E. coli, the fusion protein was also purified and proved that could induce strong cellular and humoral immune responses with appropriate adjuvant in C57BL/ 6 mice and could be used for future research.

MeSH terms

  • Adjuvants, Immunologic
  • Animals
  • Condylomata Acuminata / genetics*
  • Enzyme-Linked Immunosorbent Assay
  • Enzyme-Linked Immunospot Assay / methods
  • Escherichia coli / genetics
  • Gene Expression
  • Genetic Vectors
  • Immunity, Humoral
  • Immunization
  • Mice
  • Mice, Inbred C57BL
  • Polymerase Chain Reaction
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Recombinant Fusion Proteins / pharmacology
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics

Substances

  • Adjuvants, Immunologic
  • Recombinant Fusion Proteins
  • Recombinant Proteins