| 摘要: |
| 为了研究胸腺五肽基因的免疫增强作用，根据GenBank收录的新城疫病毒(NDV)F基因序列，设计1对特异性引物，其中上游引物含有胸腺五肽（Thymopentin，TP-5）的基因序列。以NDV Z株F基因的重组质粒为模板进行PCR扩增，扩增产物与真核表达载体pcDNA3.1/V5-His-TOPO重组，构建了真核共表达质粒pcDNATP-5NDVZF。将重组质粒pcDNATP-5NDVZF、pcDNANDVZF分别以100 μg/只的剂量免疫小鼠，用间接ELISA法和MTT法检测其免疫原性。结果表明，重组质粒均能在小鼠体内诱导相应的体液免疫和细胞免疫，其中pcDNATP-5NDVMZF的作用较pcDNANDVZF更为显著。说明共表达胸腺五肽基因可增强机体对DNA疫苗的免疫反应。 |
| 关键词: 新城疫病毒F基因 pcDNATP-5NDVZF 免疫原性 |
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| 基金项目:山东省医药卫生科技发展计划重点项目(2005ZD05)；山东省自然科学基金项目(Y2005C21) |
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| Construction of eukaryotic expression plasmid of NDV F and thymopentin gene and detection of the immunogenicity |
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| Abstract: |
| To study the immunopotentiation of thymopentin,according to the F gene sequence of newcastle disease virus published by GenBank,a pair of specific primers were designed,with up stream primers including the gene of TP-5.The NDV F gene DNA fragment was obtained by polymerase chain reaction(PCR) by using the recombination plasmid of NDVZF gene as the template.The PCR products were recombined into pcDNA3.1/V5-His-TOPO expression vector to construct eukaryotic expression plasmid pcDNATP-5NDVZF .The pcDNATP-5NDV ZF and pcDNANDVZF were separately injected into muscles of mice at a dose of 100 μg.The vaccinal immunogenicity was detected with indirect ELISA and MTT.The results showed that the recombination plasmid could induce humoral and cellular immunity.And the effect of pcDNATP-5NDVZF was more significant than pcDNANDVZF .This study indicated that the expression products of TP-5 gene could strengthen the immunogenicity of DNA vaccines. |
| Key words: newcastle disease virus F gene pcDNATP-5NDVZF immunogenicity |
