摘要
将荣昌猪外周血淋巴细胞在刀豆素A（ConA）的刺激下培养24—70h后，提取总RNA，应用RT—PCR技术扩增白细胞介素-2（IL-2）和白细胞介素-4（IL-4）cDNA，克隆到pMD18-T载体并测序。结果表明：克隆的IL-2cDNA全长为522个碱基，ORF为465个碱基，编码154个氨基酸。与GenBank公布的猪IL-2比对同源性均为99．8％；克隆的IL-4cDNA全长411个碱基，ORF为402个碱基，编码133个氨基酸，与GenBank公布的猪IL-4比对同源性均在98．0％以上，从而证实成功克隆了荣昌猪IL-2和IL-4基因cDNA。
The interleukin-2 and interleukin-4 cDNA of Rongchang Pig were amplified by RT PCR from total RNA extracted from blood lymphocytes, which were cultivated and stimulated with 7 μg/mL ConA in vitro for 24 -72 hours. The amplified cDNA was cloned into PMD18 - T vector. The nucletide acid sequences of cloned Rongchang Pig interleukin -2 and interleukin -4 cDNA were determined. The length of the IL -2 was 522 bp ,encoding 154 amino aeids; however, the length of the IL-4 was 411 bp,encoding 133 amino acids. By blasting the homologous sequences in GenBank databases,the sequence of Rongchang pig interleukin -2 gene from lymphocyte is 99. 8% percent identity to the interleukin -2 genes previously cloned from other porcines;however, the sequence of Rongchang pig interleukin -4 gene is more than 98. 0%
出处
《黑龙江畜牧兽医》
CAS
北大核心
2007年第1期10-12,共3页
Heilongjiang Animal Science And veterinary Medicine
基金
国家"973"项目(2004CCA01800)
关键词
荣昌猪
IL-2
IL-4
克隆
Rongchang Pig
Interleukin - 2
Interleukin - 4
cloning
