日本血吸虫中国大陆株成虫cDNA文库的免疫筛选及PCR初步鉴定

IMMUNOSCREENING AND CHARACTERIZATION OF SCHISTOSOMA JAPONICUM cDNA LIBRARY BY PCR

用成虫抗原免疫免血清筛选构建于表达载体λgt11的日本血吸虫成虫cDNA文库，再以多聚酶链反应（PCR）技术鉴定免疫筛选的阳性克隆。结果，对cDNA表达库中随机选取的约6.2×104个噬菌斑进行了筛选．初筛时共挑出52个可能的阳性斑，经两次复筛后，有25个噬菌仍呈阳性反应；以阳性噬菌斑DNA为模板，经PCR扩增均可获得一定大小的片段，从564bp到1200bp不等。这样能快速大量地筛选cDNA文库，便于高效地获得有价值的重组抗原基因克隆。

The adult worm cDNA library of Schistosoma japonicum (S j) on the gt11 phage vector was screened for the clones positive to rabbit serum raised against soluble adult worm antigen of Sj. The posltlve clones were analyzed by PCR amplification with prlmers on the vector. Fifty-two posltlve clones were obtalned by the first screenlng from 6. 2 ×10 of plaques and twenty-five of them were still positive after the second and the third screening by the serum. The prepared DNA of the twenty-five positive clones were used to amplify inserts by PCR and every clone had inserted cDNA fragment, the size varied from 564bp to 1200bp.This method can be of general use,it is easy to screen large number of clones from Sj cDNA library and get valuable recombinant antigen genes in an efficiency.