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当前位置: 首页 > 文章 > 山羊痘病毒P32基因真核表达质粒的构建及其诱导体液免疫应答效果 贵州畜牧兽医 2015,39 (5) 4-8
Position: Home > Articles > Construction and Effect of Immune Response of Induction Body Fluids of Eukaryotic Vectors Based on Goat Pox Virus P32 Gene Guizhou Journal of Animal Husbandry & Veterinary Medicine 2015,39 (5) 4-8

山羊痘病毒P32基因真核表达质粒的构建及其诱导体液免疫应答效果

作  者：
龙福;文明;陈光燕;王华平
单  位：
贵州省关岭自治县畜牧服务中心;贵州省望谟县草地畜牧发展中心;贵州大学动物科学学院
关键词：
山羊痘病毒;P32基因;真核表达质粒;构建;体液免疫应答
摘  要：
为探索山羊对山羊痘病毒P32基因免疫的应答效果,本研究从p MD-18T-P32中获取P32基因,插入真核表达质粒PVAX1,经PCR和酶切鉴定后,将PVAX1-P32肌肉注射小鼠,于不同时间段采集小鼠组织样本,检测分析重组质粒的分布动态和重组情况,并免疫注射山羊,于不同时间段采集山羊血清样本,检测分析重组质粒诱导山羊的体液免疫应答效果。结果显示:经PCR扩增与双酶切,PVAX1-P32均可得到与预期大小相一致的目的片段(969 bp);注射小鼠后7~45 d,在小鼠腿肌、心肌、肝脏、脾脏、肺脏、肾脏、脑、十二指肠中均可检测到目的基因,且未发现P32基因与宿主基因组整合现象;注射山羊15 d后,诱导山羊产生较高水平的总Ig G,低于弱毒疫苗组,但差异不显著(P>0.05);也能诱导山羊产生较高的抗P32-Ig G含量,高于弱毒疫苗组,差异显著(P<0.05)。结果表明:本研究成功构建了山羊痘病毒P32基因的重组真核表达质粒PVAX1-P32,能较长时间存在宿主组织器官内,可诱导山羊产生较好的体液免疫应答。
译  名：
Construction and Effect of Immune Response of Induction Body Fluids of Eukaryotic Vectors Based on Goat Pox Virus P32 Gene
作  者：
Long Fu;Wen Ming;Chen Gangyan;Wang Huaping;Center of Livestock in Guanlin County;College of Animal Science,Guizhou University;Center of Grassland and Livestock in Wangmo County;
关键词：
Goat Pox Virus;;P32 Gene;;Expression Plasmid;;Construction;;Humoral Immunity
摘  要：
To research the immune response of goat to gene vaccine,P32 gene of goat pox virus was amplified from p MD- 18T- P32 and inserted into the eukaryotic expression vector PVAX1,which identified with PCR and restriction enzyme digestion. The recombinant plasmids were injected to mice and used for analyze of distribution and verification of recombinant into genome of mice. And the plasmids were immunized to goat,then the sera samples of goats were collected for detection of the anti- P32 antibodies of goat. The Results showed as follows: the expected fragment( 969 bp size) were obtained from the recombinant PVAX1- P32 respectively by PCR and enzyme restriction. In 7d to 45 d after injection,the target gene was detected in leg muscle,heart,liver,spleen,lung,kidney,brain and duodenum of mice,and the integration phenomenon of the target gene into the host genome was not found. In 15 d to 45 d after injection,the higher level of total Ig G was detected in the sera of goat induced by the recombinant PVAX1- P32,but lower than that of the attenuated vaccine group,which the difference between them was not significant( P > 0. 05). The content of the anti- P32 Ig G in goat induced by PVAX1- P32 was higher than that of the attenuated vaccine group,which the difference between them was significant( P < 0. 05). These results demonstrated that the recombinant eukaryotic expression plasmid PVAX1- P32 are successively constructed and can exist within the host tissue organs for a long time and may induce the humoral immune response of goat.

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