摘要
利用表达载体λgt11构建鸡毒支原体 (MG)S6 株基因文库。根据载体中 1acZ基因插入失活的原理 ,重组子中大约 6 9%为重组噬菌体。应用MG纯化抗体筛选文库 ,得到 179个阳性克隆 ,不足重组噬菌体的 1%。随机选择一个阳性克隆 ,收集其蛋白进行Western免疫印迹 ,检测到 -38kD蛋白 ,表明在大肠杆菌中能合成具有免疫学活性的MG蛋白 。
A genomic library of Mycoplasma gallisepticum(MG) strain S6 DNA was generated by using bacteriophage lambda λgt11 as the vector.Approximately 69% of the resulting phages were found to be recombinants,based on the insertional inactivation of the lac Z gene of the vector. Screening of the library for the expression of MG protein antigens with antibodies purified by affinity chromatography,179 positive clones were found,that less than 1% of the total number of the recombinant phages.A single clone was chosen at random for comparison with a vector control by western immunoblot,revealing a polypeptide of 380.000 molecular weight in the clone profile but not the control profile that reacted with MG antibodies.The results demonstrated that the immunologically active MG protein were synthesized in E. coli,which may provided an improved means for antigen preparation for serologic diagnosis of mycoplasmosis. [
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2000年第5期453-457,共5页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
