Gao Yu-Wei, Xia Xian-Zhu, Wang Li-Gang, Liu Dan, Huang Geng
The Military Veterinary Institute of Academy of Military Medical Science of PLA, Changchun 130062, China.
Wei Sheng Wu Xue Bao. 2006 Apr;46(2):297-300.
H5N1 highly pathogenic avian influenza virus was highly pathogenic and sometimes even fatal for tigers and cats. To develop a new type of vaccine for Felidae influenza prevention, recombinant replication-competent canine adenovirus Type 2 expressing hemagglutinin gene of H5N1 subtype tiger influenza virus was constructed. A/tiger/Harbin/01/2003 (HSN1) HA gene was cloned into PVAX1. The HA expression cassette which included CMV and HA and PolyA was ligated into the E3 deletion region of pVAXdeltaE. The recombinant plasmid was named pdeltaEHA. The pdelta EHA and the pPoly2-CAV2 were digested with Nru I /Sal I, respectively. The purified Nru I/Sal I DNA fragment containing the HA expression cassette was cloned into pPoly2-CAV2 to generate the recombinant plasmid pCAV-2/HA. The recombinant genome was released from pCAV-2/HA, and was transfected into MDCK cells by Lipofectamine. The recombinant virus named CAV2/HA was gained. Anti-H5N1 influenza virus HI antibody (1:8 - 1:16) was detected in the cat immunized with CAV-2/HA.
H5N1高致病性禽流感病毒对老虎和猫具有高致病性,有时甚至是致命的。为开发一种新型的预防猫科动物流感的疫苗,构建了表达H5N1亚型老虎流感病毒血凝素基因的具有复制能力的重组犬2型腺病毒。将A/老虎/哈尔滨/01/2003(HSN1)HA基因克隆到PVAX1中。将包含CMV、HA和PolyA的HA表达盒连接到pVAXdeltaE的E3缺失区域。重组质粒命名为pdeltaEHA。分别用Nru I /Sal I消化pdelta EHA和pPoly2-CAV2。将含有HA表达盒的纯化的Nru I/Sal I DNA片段克隆到pPoly2-CAV2中,以产生重组质粒pCAV-2/HA。从pCAV-2/HA中释放重组基因组,并通过Lipofectamine转染到MDCK细胞中。获得了重组病毒CAV2/HA。在用CAV-2/HA免疫的猫中检测到抗H5N1流感病毒HI抗体(1:8 - 1:16)。