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Construction of hpv-16-l1,HSV-gD Vaccines with their protein expressions in DH5 α and possibly dcs present hpv-16-l1 protein to lymphocytes

Construction of HPV-16-L1,HSV-gD Vaccines with their protein...

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2012全国临床微生物与感染免疫学术研讨会

作者： Yan Bing~(1,2),Meng Xia~1,Jiang Yuanhong~(1,3),Wang Jianyu~1,Fan Fan Xiaoguang~1 1.Department of Microbiology and Immunology,2 Department of Electron Microscope,Shanxi Medical University 2.College of Clinical Medicine at five years length 2006,West China Center of Medical Science,Sichuan University 3.Department of Electron Microscope,Shanxi Medical University

Objective It is well known that cervical cancer has close relationship with hpv and HSV and the investigation showed that 50%cervical cancer patients had been infected by hpv 16,of which l1 is the major capsid protein, an antigen can stimulate lymphocyte in the host to produce antibody to neutralize the ***Ⅱis the cofactor in the processing and development of cervical *** order to obtain the recombinant protein of pBV220- hpv16l1 and pBV220-HSVgD,it’s necessary to induce their expression in ***5αand purify the acquired *** use these two sets of recombinant protein to prepare for the polyclonal antibodies by immunized New Zealand White rabbits. Methods The source of the DNA were extracted and amplified by PCR reaction using sense primer and antisense primer according to the hpv type 16-l1 gene is the tissue of patients,whose diagnose not only confirmed clinically,but also reassured by their tissue sample ***,gD gene was also amplified from HSV as well. The *** DH5αwith the recombinant plasmids express pBV220-hpvl1 and pBV220-HSVgD respectively in the form of inclusion body after induced at 42℃for 5 hours or 6 hours *** obtained protein can be detected by *** cracking the bacteria with sonar,we got the inclusion bodies that contain the target *** they were washed in solution that contains 2 mol/l urea as final concentration for acquire the recombinant proteins at high purity,following by the Branford method for *** we immunize the rabbit with these proteins by injecting 1 mg/kg for the first inoculation and 0.5 mg/kg for the rest 6 shoots or till the titer and specificity of the hpvl1 and HSV-gD antibodies were high enough to be detected by agarose double immune *** the blood of the rabbits from their hearts were collected and sera were kept in -20℃environment for the western blot. Recombinant hpv16-l1 protein was presented by Dendritic Cells(dcs) to lymph

关键词： Constructs of HSV -gD and hpv -16-l1 Recombinant protein expression Polyclonal antibody dcs present hpv-16- l1 protein to lymphocytes

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