山西本地株人乳头瘤病毒16-L1和单纯疱疹病毒-gD预防性重组疫苗的构建及筛选  

The constructs and Screening of Shanxi Local strain of HPV-16-L1 and the HSV-gD recombinant prophylactic Vaccines

作　　者：阎冰[1] 孟霞[1] 阎来喜[1] 王建宇[1] 郭松佳[1] 樊晓光[1] Yan Bing;Meng Xia;Yan Laixi;Wang Jianyu;Guo Songjia;Fan Xiaoguang(Laboratory of Reproductive Immunology,Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学生殖免疫研究室,太原030001

出　　处：《中国药物与临床》2021年第17期2929-2933,共5页Chinese Remedies & Clinics

摘　　要：目的子宫颈癌是在全球范围内女性第二常见的恶性肿瘤。基于山西人人乳头瘤病毒(HPV)16-L1当地病毒株的基因和单纯疱疹病毒(HSV)-gD基因,构建了HPV 16-L1和HSV-gD重组预防疫苗,以利于预防山西省妇女宫颈癌。方法选取山西省妇女宫颈癌患者的组织,提取了患者基因组DNA后,依HPV 16-L1基因型的正向引物和反向引物,通过聚合酶链反应(PCR)扩增获得HPV 16-L1基因,并从HSV病毒株中PCR扩增gD基因。将这2个已验证的基因克隆到pMD18T中,且分别插入pcDNA3.0和pBV220质粒内,通过限制性内切酶和DNA测序等方法,构建了此2个重组基因,并验证了2个重组基因的正确性。结果获得了1.59kb HPV 16-L1基因和311bp HSV-gD片段基因,及其分别在pcDNA3.0和pBV220中的表达产物。L1基因和gD基因的序列与我们设计的序列匹配。结论实验的关键是扩增了中国山西地区携带HPV患者体内的HPV 16型病毒L1全长基因。建立了HPV 16-L1和HSV-gD质粒的构建体,为在动物和临床试验中,不久的将来,应用宫颈癌预防性疫苗,提供了第一步原核细胞生物和真核细胞生物方面的研究。Objective Carcinoma of the cervix is the second most frequent malignancy in women on a global scale.The construction of HPV 16-L1 and HSV-gD recombinant prophylactic vaccine has been made based on the genes of Shanxi local strain of HPV 16-L1 and HSV-gD in order to prevent cervical cancer in women in Shanxi province.Methods The source of the human genomic DNA that were extracted and amplified by PCR reaction using sense primer and antisense primer according to the HPV type 16-L1 gene which is in the tissue of cervical cancer patients and gD gene was amplified from the strain of HSV as well.The two verified genes were cloned into pMD18T,then inserted into pcDNA3.0 and pBV220 respectively,which results were verified by their constructs through restricted endogenous enzyme cut and DNA sequencing.Results The 1.59kb HPV 16-L1 gene and 311bp HSV-gD fragment were obtained.The proteins expresssion of L1 gene and gD and which gene matched with the ones we designed in the hypothesis.Conclussion The critical experiment was amplified the gene of HPV type 16-L1 by PCR in patients who carried HPV in Shanxi area China.The manipulated the constructs of HPV 16-L1 and HSV-gD in plasmids provide the first step towards usage of expression of prokaryote and eukaryote of prophylactic vaccine of cervical cancer in animal and clinical trial in the near future.

关 键 词：宫颈肿瘤 人乳头瘤病毒16 单纯疱疹病毒疫苗 

分 类 号：R737.33[医药卫生—肿瘤]