华北农学报2011，Vol.26Issue(5)：71-75,5.
猪白细胞介素-2基因的克隆及其重组禽痘病毒载体的构建
Cloning of Porcine IL-2 Gene and Its Construction of Recombinant Fowlpox Viruses Vector
摘要
Abstract
Porcine IL-2 gene was amplified by RT-PCR from the total RNA extracted from the splenocyte of Yu' nan pig. The PCR product was cloned and sequenced. The porcine IL-2 (pIL-2) gene was digested by Bamli I and cloned into pSY538 vector which was digested by Bamli I and dephosphorized by calf alkaline phosphatase. By means of the PCR .restriction enzyme digestion and sequencing, the recombinant plasmid pSY538/pIL-2 was selected. After pSY538/pIL-2 was digested by Not I ,the pIL-2 fragment containing LP2 and EP2 was achieved and sub-cloned into pSY681 vecter containing LacZ gene,then the recombinant plasmid pSY681/pIL-2 was developed. The results indicated that the cloned plL-2 gene had 482 bp, including one ORF(465 bp). The IL-2 gene sequence of Yu'nan pig had 99. 9% nucleotide homology with the 5 pig IL-2 genes published in GenBank. Enzyme digestion and DNA sequencing results confirmed that the sequence of the recombinant plasmid pSY681/pIL-2 contained the target fragment,and the ligation part was correct. The results showed that the recombinant plasmid pSY681/pIL-2 was constructed correctly, paving the way for further study of biological function and further application of porcine IL-2.关键词
猪白细胞介素-2/克隆/重组禽痘病毒/载体/构建Key words
Porcine IL-2/Clone/Recombinant fowlpox virus/Vector/Construction分类
农业科技引用本文复制引用
王淑娟,陈红英,魏战勇,刘金朋,耿静微,崔保安..猪白细胞介素-2基因的克隆及其重组禽痘病毒载体的构建[J].华北农学报,2011,26(5):71-75,5.基金项目
国家"十一五"科技支撑计划专项(2006BAD06A08) （2006BAD06A08）
