摘要
克隆牦牛白细胞介素-4(IL~4)基因,并对其进行遗传演化分析。从刀豆素(ConA)和脂多糖(LPS)联合刺激培养的牦牛外周血淋巴细胞提取总RNA.利用RT—PCR方法扩增牦牛IL-4全长cDNA,将其克隆到pMD18~T载体上,测序后进行序列分析。成功克隆到牦牛IL-4基因全序列,序列分析表明,克隆的牦牛IL-4基因序列与GenBank所登录的牛IL-4核苷酸序列及推导的氨基酸序列的同源性分别这99.8%和100%,与人、猕猴、猪、山羊、马等物种的核苷酸及其推导氨基酸序列的同源性分别在44.4%~96.3%和27.4%~91.1%之间。在国内首次成功地从牦牛外周血淋巴细胞中克隆到IL-4的基因,其ORF为408bp,推导编码135个氨基酸。
Cloning and genetic analysis of yak interleukin-4 gene were conducted. The full length cDNA of boiL-4 was amplified by RT-PCR from the yak peripheral blood lymphocyte stimulated by ConA and LPS, and was cloned in pMD18-T vector and sequenced. Homologies of nucleotide and deduced amino acid sequences between the yak IL-4 and registered boiL4 in the GenBank are 99.8% and 100%,respectively. Homologies of yak IL-4 nucleotide sequence compared with Homo, Macaca, Sus scrofa, Capra and Equus are in 44.4%-99.8%. Homologies of amino acid sequence of them are in 27.4%-91.1%. In conclusion, IL- 4 containing an ORF of 408 bp and coding 135 amino acids was first cloned successfully from the yak PBMC in China.
出处
《动物医学进展》
CSCD
2008年第8期24-28,共5页
Progress In Veterinary Medicine
基金
国家高新技术"863"项目(2006AA10A203)
甘肃省自然基金项目(3ZS061-A25-081)
关键词
白介素4
克隆
序列分析
牦牛
interleukin-4
cloning
sequence analysis
yak