Gene Cloning And Expression Of HPV Type 6b L1 Protein In Eukaryotic Cells.

Objectives To construct PEGFP-HPV6bL1 expression vector and to detect its transient expression in eukaryotic cells,which might have great potential for genetic engineering vaccine against condylome acuminatum(CA) and provide useful dates in stucying the biology characteristic of HPV L1 protein.Methods The gene fragment of HPV6bL1 has been amplified by PCR and cloned into the vector PEGFP-C1 Then the recombinant expression vector was transformed into coli DH5a,which was identified by BamHI and HandIII digestion. The positive vector was sequenced to get late gene L1 recombinant sequence.Then the recombinant was transfected into COS-7 cells by techniques of gene transfection .the vector expressed the gene HPV6bL1 in COS-7 cells ,which was detected by fluoroscopy and RT-PCR.Results Identification of PEGFP-HPV6bL1 by enzyme digestion and sequencing showed that the length,inserted location and direction of target which was inserted into the recombinant was correct and the expression of EGFP in transfected cell was observed.Conclusions The PEGFP- HPV6bL1 expression vector makes it easy to assess theexpression of PEGFP- HPV6bL fusion protein and to sift the transfected cells.It is quite potential in the future research for the development of genetic engineering vaccine.