Zheng L, Liu S G, Cheng S Q, Xu H
Department of Biotechnology, Sichuan University, Chengdu.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1993;11(4):258-61.
This paper reported the immunoscreening of three recombinants expressing secretory antigens from the genomic DNA library of F. hepatica and the primary study of expression of the three recombinants. To construct the genomic DNA library, DNA from the adult F. hepatica was cut with Sau3AI to an average length of about 2.0 kb and inserted into the BamHI site of the expression vector pUC18, the size of the genomic DNA library of F. hepatica being 1.5 x 10(5) recombinants. Three recombinants expressing antigenic determinants of F. hepatica pFH16, pFH23, pFH48 were detected through primary screening and rescreening with F. hepatica infected rabbit serum (1:50) preabsorbed to remove antibodies to E. coli and SPA-HRP (1:40). The test of the ability of expressing fusion proteins showed pFH16 > pFH48 > pFH23. These studies provide the possibility of further research on the expression of recombinant antigenic genes, the immunity of their expressed products and the protection of animals.
本文报道了对三种表达肝片吸虫基因组DNA文库分泌抗原的重组体的免疫筛选及其表达的初步研究。为构建基因组DNA文库，用Sau3AI切割肝片吸虫成虫的DNA，使其平均长度约为2.0 kb，然后插入表达载体pUC18的BamHI位点，肝片吸虫基因组DNA文库的大小为1.5×10⁵个重组体。通过用预先吸收以去除抗大肠杆菌抗体的肝片吸虫感染兔血清（1:50）和SPA-HRP（1:40）进行初次筛选和复筛，检测到三种表达肝片吸虫抗原决定簇的重组体pFH16、pFH23、pFH48。融合蛋白表达能力检测显示pFH16>pFH48>pFH23。这些研究为进一步研究重组抗原基因的表达、其表达产物的免疫性及动物保护提供了可能性。
