鸡新城疫系疫苗诱导鸡IL-2基因转录及cDNA克隆的研究

Transcription and cDNA cloning of chicken interleukin-2 induced chicken embryo splenocytes by Newcastle Disease viruse

应用RT-PCR方法从鸡新城疫 系疫苗接种鸡胚的脾淋巴细胞中扩增出IL-2基因cDNA。结果表明,以鸡新城疫 系疫苗稀释50倍或100倍接种10日龄鸡胚,培养48h的脾脏提取mRNA的扩增效果最好。测序结果显示,所扩增片段的核苷酸序列及其所编码的氨基酸序列,与已发表的IL-2基因的同源性分别为97.7%～99.8%和95.1%～99.3%。其中第8位、68位和130位氨基酸(分别是L,V和S)的变异是其他品种鸡所没有的。

A cDNA encoding chicken interleukin-2 (chIL-2) was cloned from Luoyang local chicken embryo splenocytes by reverse transcription-polymerase chain reaction (RT-PCR).The results showed that the best factor for mRNA isolated from spleen cells of chicken embryo was 48 h after being inoculated with 50× or 100× diluted Newcastle Disease virues.Sequence analysis showed that the chIL-2 cDNA was 432 bp and encoded a protein 143 amino acids compared with reported chIL-2.The homology of nucleotide acid and amino acid were 97.7%-99.8% and 95.1%-99.3%,respectively.Especially the amino acids in sites 8,68 and 130 (L,V and S) were different from other strains.These provide a basis for the study of the structure and function of chIL-2 gene.