GPMV NA-1株致弱F基因真核质粒的构建及免疫试验  

Construction and immunization of eukaryotic plasmid of attenuated fusion gene of GPMV NA-1 strain

作　　者：李国江[1,2] 张立春[1] 张秀峰[1] 姜成[1] 刘立明[1] 丁壮[2] 

机构地区：[1]吉林农业科技学院,吉林吉林132101 [2]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《中国兽医学报》2010年第11期1427-1431,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30771606)

摘　　要：将本实验室致弱的GPMV F基因克隆到真核表达载体pCI-neo中,构建pCI-rf质粒,经脂质体法转染BHK细胞后,Western blotting检测pCI-rf能正常表达。采用pCI-rf和本实验室构建的含有GPMV的F基因的真核表达质粒pCI-f分别肌注7~9日龄雏鸡,间接ELISA检测抗体,并采用淋巴细胞增殖试验和特异性CTL杀伤活性试验对试验鸡的细胞免疫进行检测,最后做攻毒保护试验。结果表明,pCI-f和pCI-rf能诱导试验鸡产生细胞免疫和体液免疫应答,与LaSota疫苗比较,免疫效果没有显著差异。PCR扩增检测结果显示,未能从免疫雏鸡各组织的基因组中扩增出F基因片段。The attenuated GPMV F gene was cloned into the eukaryotic expression vector of pCI-neo and the pCI-rf plasmid was obtained.The pCI-rf plasmid was transfected into BHK cells and the fusion protein was detected through Western blotting successfully.The 7 to 9-day-old chicken was immunized intramuscularly with pCI-rf and pCI-f separately.The antibody level was detected by indirect ELISA and the cell immunity was detected by lymphocyte proliferation assay and specific lysis activity of CTL test.The immunized chicken was challeged finally.The result showed that both of pCI-rf and pCI-f induced cell immunity and humoral immunity responses and the difference of immune effect was not obvious comparing with Lasota vaccine.The PCR result showed that F gene fragment was not detected from the genome of immunized chicken tissues.

关 键 词：鹅副黏病毒 致弱F基因 核酸疫苗 免疫原性 

分 类 号：S852.65[农业科学—基础兽医学]