Zhang W, Zhang J, Liu S
Institute of Parasitic Diseases, Chinese Academy of Preventive Medicine, Shanghai 200025.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1999;17(2):97-100.
To study the association expression of two genes encoding GST of Schistosoma japonicum and K99 of enterotoxigenic Escherichia coli.
PCR technique was used to gain the K99 gene. After digestion with BamH I and EcoR I, the gene was cloned into the plasmid vector pUC18 by using recombinant DNA techniques. The other target gene of GST in pSj5 plasmid was obtained by EcoR I digestion, and was then ligated into the recombinant plasmid pUC18-K99. The expressed product was assayed by SDS-PAGE, Western blotting, reversal IHA, ELISA and transmission electron microscopy.
Co-expression product of around 50 kDa was obtained. The protein was recognized by anti-GST and anti-K99 antibodies (ELISA and IHA), and acted as pilus on the surface of transformed DH5 alpha E. coli bacteria.
Co-expression of the genes encoding GST and K99 was successfully achieved.
研究日本血吸虫谷胱甘肽S-转移酶（GST）两个编码基因与产肠毒素大肠杆菌K99的联合表达。
采用PCR技术获取K99基因。经BamH I和EcoR I酶切后，利用重组DNA技术将该基因克隆至质粒载体pUC18。通过EcoR I酶切从pSj5质粒中获取GST的另一个靶基因，然后将其连接至重组质粒pUC18-K99。采用SDS-PAGE、Western印迹、反向间接血凝试验（IHA）、酶联免疫吸附测定（ELISA）及透射电子显微镜对表达产物进行检测。
获得了约50 kDa的联合表达产物。该蛋白可被抗GST和抗K99抗体识别（ELISA和IHA），并在转化的DH5α大肠杆菌表面起菌毛作用。
成功实现了编码GST和K99基因的联合表达。
