【摘要】 用质粒表达载体pSPORT 1构建了猪囊尾蚴cDNA文库。将已构建好的cDNA表达文库用SOC按一定比例稀释后 ,均匀涂布于LB板 (Amp+ )上 ,37℃培养箱中倒置培养过夜。将生长良好的菌落转印于NC膜 ,并将此膜置于加有IPTG的LB板上 ,37℃诱导培养 4h。取下NC膜 ,在氯仿蒸气中熏蒸 15min后 ,在裂解液中过夜裂解。经洗膜、封闭 ,加人囊尾蚴阳性血清 ;再洗膜、封闭 ,加羊抗人IgG HRP ,与底物作用后 ,初步筛选得到了阳性克隆。对可疑阳性克隆 ,用同样方法反复筛选 ,直到完全确定为止更多还原

【Abstract】 The cDNA library from Cysticercus cellulosae had been constructed with the vector (pSPORT-1). After the library was diluted with the SOC and distributed well on the LB plates(Amp~+),(the bacteria) had been cultured for 14 hours. While the bacteria were grown well and transferred on the (nitrocellulose) filters(NC) and then put the NC on the LB plates(containing IPTG),the bacteria had been cultured for 4 hours at 37 ℃.The NC were stifling in chloroform for 15 min and put into lysis-buffer for 14 hours. After the NC were washed and closed, then put into the positive serum. If sheep anti-people IgG-HRP and substrate were added, the positive clones would appear.更多还原