表达禽流感病毒核蛋白重组噬菌体的构建

摘要：利用PCR技术，扩增去除部分序列、长度为1320 bp、编码440个氨基酸的禽流感病毒(AIV)NP基因片段，将其克隆至pR质粒的T4噬菌体SOC基因C末端获得重组质粒pR-NP，以此重组载体转化大肠杆菌E2，用溶菌酶缺陷噬菌体T4-Z1感染重组E2菌，重组载体与缺陷噬菌体T4-Z1基因发生同源重组，将NP基因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-NP。经Western blot检测证实，T4-Z1-NP表达的NP融合蛋白具有免疫学活性。成功构建了表达禽流感病毒核蛋白的重组噬菌体。

Construction of Recombinant Bacteriophage T4 Expressing np Gene of Avian influenza virus

ABSTRACT：The NP gene segment deleted part of sequence，with a length of 1320bp，and coding for 440 amino acids，was amplified by PCR．The NP gene fragment was then inserted into the EcoR I site of pR vector to obtain recombinant plasmid pR-NP. The recombinant plasmid was used to transform E．coli E2, the host bacteria of T4 phage. and then infected with lysozyme-defective phage T4-Z1.The NP gene were then integrated into the SOC site of the genome of SOC-deleted T4 phage mutant by homologous recombination. The recombinant phage were screened by PCR and designated as T4-ZI-NP．The immunological test applying Western blot showed that SOC-NP fusion protein expressed by the phage T4-ZI-NP could react to AIV specifically. Recombinant bacteriophage T4 expressing the NP Gene of Avian Influenza Virus was constructed successfully.