摘要
本文以鸡外周血单个核细胞总 Ｒ Ｎ Ａ 为模板，据已报道的８ 种哺乳动物 Ｉ Ｌ—１β核酸序列保守区设计合成１ 对引物，反转录合成ｃ Ｄ Ｎ Ａ 链，经 Ｐ Ｃ Ｒ 扩增，回收扩增后的 Ｄ Ｎ Ａ 片段，用ｋｌｅｎｏｗ 酶处理，与 Ｐｕ ｃ１ ９ ／ Ｓｍ ａ Ｉ 连接构成重组质粒，转化宿主菌大肠杆菌 Ｊ Ｍ１ ０ ９ ，筛选出含有插入片段的重组质粒，用 Ｐ Ｃ Ｒ 及酶切分析鉴定，结果表明获得了带有鸡 Ｉ Ｌ－ １β片段的ｃ Ｄ Ｎ Ａ 克隆；再经序列分析测定，得知此片段长为２７０ｂｐ ，与人及小鼠 Ｉ Ｌ—１β基因核苷酸序列同源性分别为４５ ％ 和３０ ％ 。
A pair of primers were designed according to the described interleukin-1β sequence of mammals,such as human,murine,equine,bovine,ovine,rabbit.And the total RNA in peripheral blood mononuclear cell (PBMC)were produced by stimulating chickens pripheral blood with Lipopolysaccharide(LPS).The total RNA in PBMC were used as sample to syntheside cDNA through RT.About 0.27kb fragment which were correspord to be predicted were amplified.Analysis of the chicken IL-1β cDNA sequences showed that 45%and 30% homology identity to the corresponding human,murine sequences was 45% and 30% respectively.
基金
国家自然科学基金
