Experssion And Identification Of Human Papillomavirus Type 58L1 Gene In Picha Pastoris

The human papilloma virus is can cause the human skin, the mucous membrane papilloma or a wart kind of tumor related virus. At present, HPV has separated 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.The recent years literature reported and the epidemiology research indicated, has the high carcinogenicity risky besides HPV16 and 18, but also some 31, 33, 45 and 58. International studies have consistently showed a high prevalence of HPV infection among women with invasive cervical cancer and that HPV DNA can be detected in approximately 90% of al1 cervicalcancers. Recently, strong evidence from epidemiologicalsurveysand molecular biological studies has implicated thathuman papiL1omavirus type 58(HPV58) infection is closelyrelated with cervical cancer. Particularly, in SouthernChina, thepositive rate of HPV58 was the higher in biopsies of cervical cancer. The positive rate of HPV58 increased gradual1y with the malignant extent of the lesions assayed, which is the same as that of HPV16. HPV58 has been shown to be one of the most important highly-oncogenic, risky HPV types in China.At present, cervical cancer oneself becomes endangers one of woman health main malignant tumors, the disease incidence rate has the tendency which advances year by year, but because HPV still could not pass through the tissue culture until now 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, domestic and foreign to HPV16, 18 has done the massive work, but to the HPV58 research, specially the HPV58 structural gene constructs, the expression and the genetic engineering vaccine research are extremely 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.We first take the HPV58 gene group as the template,obtains The L1 Major capsid protein using the PCR method and constructs to the secretion yeast expresses carrier pPICZαB forms the conformity material particle, The electric shock transforms the GS115 yeast cell, the screeningmasculine gender reorganization, A selection expression quantity higher strain carries on the 5L fermentation pot the expression, Expresses the product after the purification, carries on the Western-Blot appraisal. The result showed that, is clear on the fermentation fluid has thespecial protein belt, about molecular weight 56kD, The product after the purification, the immune body examination which prepares by this experiment, proved we obtained the goal protein. This study provides a good basis for identification of the biological function and preparation of HPV58L1 specific antibody and VLPs assembly and the preparation corresponding immune body from now on using the L1 protein, then developed the HPV58 genetic engineering vaccine and the HPV multi-valence vaccine has laid the good foundation and the condition.