摘要
用PCR方法扩增不含信号序列、长度为1566bp、编码522个氨基酸的新城疫病毒(NDV)F基因片段,将其克隆至pR质粒获得重组质粒pR-F,以此转化大肠杆菌E2,用溶菌酶缺陷噬菌体T4-Z1感染重组E2菌,F基因经同源重组整合到噬菌体基因组中,用PCR方法筛选重组噬菌体并命名为T4-Z1-F。将T4-Z1-F制备成油乳剂疫苗免疫10日龄SPF鸡,25日龄加强免疫1次。结果表明,表达F蛋白的重组噬菌体具有良好的免疫原性,可诱导免疫鸡产生特异性抗体并对NDV强毒攻击提供部分免疫保护。
The F gene segment deleted the F0 signal sequence, with a length of 1566bp, and coding for 522 amino acids, was amplified by PCR. The F gene fragment was then inserted into the EcoRⅠsite of pR vector to obtain recombinant plasmid pR-F. The recombinant plasmid was used to transform E.coli E2, the host bacteria of T4 phage, and infected with lysozyme-defective phage T4-Z1. The F gene was integrated into genome of T4-Z1 by homogenous recombination. The recombinant phage were screened by PCR and designated as T4-Z1-F. Oil-emulsion vaccine was prepared with concentrated phage T4-Z1-F and used to inoculate 10-day-old specific-pathogen-free(SPF) chickens, all birds were given a booster at the age of 25-days. The result showed that the recombinant F protein was highly immunogenic, and was able to induce specific antibody against NDV to partially withstand virulent NDV challenge.
出处
《中国农业科学》
CAS
CSCD
北大核心
2005年第6期1270-1274,共5页
Scientia Agricultura Sinica
基金
国家"863"计划资助项目(2002AA245051)
