PRELIMINARY STUDY OF A NOVEL HUMAN PAPILLOMAVIRUS TYPE 16L1／E6-E7 CHIMERIC RECOMBINANT DNA VACCINE

Objective Preparations of HPV16 L1/E6 and L1/E7 prophylactic and therapeutic DNA vaccines. Methods The nucleotides within HPV16 E6 and E7 genes, which are responsible for viral transforming activity, were mutated by mage primer site-directed mutagenesis method. The correctly mutated E6 and E7 fragments were separately cloned into an eukaryotic expression vector pVAX1, together with HPV16 L1 gene, generating chimeric recombinants plasmids 1MpVAX1-L1E6, 2MpVAX1-L1E6, 1MpVAX1-L1E7, 2MpVAX1-L1E7 and 3MpVAX1-L1E7. CHO cells were transiently transfected with the individual DNA vaccines by calcium phosphate method. Target protein expressions in the extracts of the transfected cell lines were measured by ELISA and immunohistochemistry, with HPV16 L1 and E6 specific monoclonal antibodies. Results ELISA assays showed the P/N ratios in the cell extracts transfected with L1E6 and L1E7 plasmids were more than 2.1. Immunohistochemistry revealed brownish precipitant signal in cytoplasm and nuclei of the transfected cells. Conclusion Successful constructions of prophylactic and therapeutic DNA vaccine plasmids lay solid foundation for future animal experiment and clinical trial.     

非常任务

汽车比赛中,各种作用的工作车成为保证比赛顺利进行的重要组成部分。拉力赛中．有比赛开始之前各种作用的开路车和比赛结束之后的收尾车,场地比赛中,最惹眼的是比赛之前和事故发生之后出动的安全车。与上述两项比赛相比,场地越野比赛的节奏比较慢、车辆通过各种障碍时．具有极强的表演性．就是出现失误,也极具感染力。     

K30试验法在大连新港油罐工程中的应用

通过大连港新港2座5万m^3油罐K30试验检测实例，介绍K30试验的具体方法，从理论上阐明了K30值的物理意义。