Study On Self-assembly Of Human Papilloma Virus Type 58 L1&L2 Protein Into Virus-like Particles

The human papilloma virus can cause the human skin cancer, the mucous membrane papilloma and a wart kind of tumor. At present, HPV has been identified more than 100 types. Among them, infection genital tract HPV acts according to its pathogenicity to be possible to divide into the highly risky types and the low risky types. The low risky types mainly includes 1, 2, 6, 11 and so on, mainly causes the benign lump or the wart, like flat wart and incisive condyloma; The high risky types mainly includes 16, 18 and so on, mainly causes the skin mucous membrane malignant tumor, like cervical cancer and so on.At present, cervical cancer has become one of the most dangerous malignant tumors cancers which endangers woman's health, the disease's incidence rate has increased year by year, but because HPV still could not pass through the tissue culture to carry on the multiplication, also the viral nucleic acid included the latent carcinogenicity, therefore was unable to prepare the natural vaccine to prevent. On clinical high risk type of HPV infects the easy canceration, in front of the cancer the pathological change to use the medicine treatment result to be bad, spends high; To the middle or later period cervical cancer of chemotherapy, the surgery treat is not ideal. Therefore, studies and develops the HPV high risky types genetic engineering vaccine watershed.HPV prophylactic vaccine research oneself receives the domestic and foreign medical arenas universal attention. Chooses the ideal goal gene to develop the genetic engineering vaccine to be extremely important. The HPV genetic engineering vaccine goal gene choice mainly concentrates in early gene E6, E7 and later period gene L1, in the L2 gene.The therapy vaccine often applies E6, the E7 early gene, but the preventive vaccine often applies L1, the L2 gene. The HPV later period gene area codes two structure proteins: Main capsid protein L1, the structure is conservative, is the main kind of specificity antigen; minor capsid protein L2, is the specificity antigen. The L1 Major capsid protein has the conservative nature as well as with the natural viral pellet similar antigen epi-position, therefore can be induced in the protection and the immune body, may prevent HPV the infection. Therefore, the HPV prophylactic vaccine applied research mainly concentrates at present on HPV outer, covering protein L1.At present, according to the recent literatures and the epidemiology researches,it is indicated that HPV58 has become one of the most important highly-oncogenic, risky HPV types as the HPV16 in China.There are many domestic and foreign researches has been done to HPV16,and HPV 18,but the researches to the HPV58 specially to the HPV58 structural gene constructs, the expression and the genetic engineering vaccine , are still few. Based on HPV58 in the our country woman cervical cancer special position, as well as the clinical treatment faces all sorts of difficulties, the research develops the HPV58 preventive vaccine, then finally develops the HPV high risky type the union vaccine, to prevented HPV infects and the malignant tumor which causes by it has the significant practical significance.In our research, we use the vectors of pRSET-BHPV58L1 and pRSET-BHPV58L2 which have been constructed ,and the BL21 E. coli which could be induced by IPTG to express the protein of HPV58L1 and the protein of HPV58L1 . And then we purify these two proteins and renature HPV58L1 protein which is denaturalized during the purification process ,then we obtained VLP consist only by the protein of HPV58L1,.After that we try to assemble these two proteins into VLP in vitro.According to the result of WsternBlot measure ,we finally get the HPV58VLP but there is only HPV58L1 protein involved.This study provides a good basis for the research of HPV58 specific antibody and the VLPs assembly .It is helpful for the development of immune body which fight against HPV using the L1 protein, and also build a solid foundation for developing the HPV58 genetic engineering vaccine and the HPV multi-valence vaccine .