摘要
[目的]为进一步研发新型抗病毒制剂和疫苗佐剂、有效控制猪重大传染病奠定基础。[方法]根据GenBank中猪白细胞介素-18基因(IL-18)序列,设计1对特异性引物,应用RT-PCR方法从河南三元杂交猪脾脏淋巴细胞中扩增猪IL-18基因,并进行克隆、序列测定和分析。[结果]猪IL-18基因全长为579 bp,编码192个氨基酸的无活性前体蛋白,但前体蛋白并不含典型的疏水信号肽,在第35位氨基酸残基处有1个潜在的白细胞介素1β转换酶(ICE)剪切位点,剪切后变为具有生物活性的猪IL-18成熟蛋白。序列分析结果表明,该试验获得的猪IL-18基因与已知猪IL-18基因序列同源性很高,均为96%以上,其推导的氨基酸同源性在98%以上。[结论]该研究成功构建了猪IL-18基因的重组真核表达载体并得到了具有生物活性的瞬时表达产物。
[ Objective] The study aimed to lay a foundation for the development of new type antiviral agents and vaccine adjuvants so as to control the severe porcine infectious diseases effectively. [ Method] According to the porcine intedeukin-18 gene （IL-18） published in Gen- Bank, a pair of specific primers was designed for amplifying the porcine IL-18 gene from the spleen lymphocytes of Henan three-way crossbreeding pigs by RT-PCR. Subsequently, the gene was cloned, sequenced and analyzed. [ Result] The porcine IL-18 gene was 579 bp, encoding an inactive precursor protein with 192 amino acids. The precursor protein had no typical hydrophobic signal peptide and contained an unusual leader sequence composed of 35 amino acids that was cleaved for biological activity by Interleukin-1 β converting enzyme （ICE）. After comparing the IL-18 gene obtained in this experiment with other known porcine IL-18 genes, it showed that the nucleotide sequence homology was over 96% and the deduced amino acid homology was more than 98%. [ Conclusion] The eukaryotic expression recombinant vector for por- cine IL-18 gene was constructed successfully, which could instantaneously express products with biological activity.
出处
《安徽农业科学》
CAS
北大核心
2009年第6期2413-2415,共3页
Journal of Anhui Agricultural Sciences
基金
河南省基础与前沿技术研究计划项目(072300430060)
河南省重点科技攻关项目(072102130023)
