人乳头瘤病毒16型E7核酸疫苗的构建及鉴定

Construction and Identification of HPV16E7 Nucleotide Vaccine

人乳头瘤病毒16 型在引起宫颈上皮转化过程中, 持续表达E7 蛋白, 有可能成为肿瘤特异性移植抗原。本文将E7蛋白的编码基因定向克隆于真核表达载体构建了HPV16E7HB核酸疫苗, 将该疫苗直接注射BALB/c 小鼠和Wistar 大鼠股四头肌, 动态观察, 28d 后仍能测出E7DNA 的扩增。提取肌组织RNA, RTPCR检测老鼠特异性E7 的转录情况, 3/3 大鼠,1/10 小鼠为阳性; 用ELISA法在免疫小鼠血清中检出了特异性抗E7 抗体。证明, HPV16E7HB

Human papilloma virus type 16 persistently expresses E7 protein during transformation of cervical epithelial cells,and the E7 protein may act as a tumor specific transforming antigen We had constructed a HPV16E7 HB nucleotide vaccine,and this vaccine was injected directly in to the quadriceps muscles of BALB/c and Wistar mice The results of dynamic observations showed that amplication of E7 DNA could be detected after 28 days of inoculations After extracting the RNA from muscle tissues and determing the transcription of specific E7 by RT PCR,it was found that the positive rates of RT PCR of BALB/c and Wistar mice were 1/10 and 3/3 respectively By using ELISA method,the E7 specific antibodies could be detected in the sera of immunized mice These results demonstrated the successive construction and effective expression of the related genes in vivo and in vitro