摘要
以伴刀豆球蛋白A(ConA)诱导的猪外周血淋巴细胞中提取的总RNA为模板,采用RT-PCR技术扩增出约500 bp的DNA片段,对阳性克隆进行测序与分析。结果:所克隆的基因与GenBank上公布的猪白细胞介素2(PoIL-2)基因的同源性为100%,表明试验成功获得了PoIL-2基因的全序列克隆;以该重组质粒为模板进行PCR,扩增出PoIL-2成熟蛋白的基因片段,连接真核表达载体pPICZαA,成功地构建了重组PoIL-2成熟蛋白基因的真核表达载体pPICZαA-PoIL-2;电转化pPICZαA-PoIL-2于巴斯德毕赤酵母X-33,诱导表达后进行表达产物的SDS-PAGE鉴定,结果表明试验成功地建立了重组PoIL-2的酵母表达系统。
The total RNA extracted from the peripheral blood of porcine stimulated with concanavalin A was extracted. It was amplified by RT - PCR. The positive clone were sequenced. The result indicated that the cloned gene had the identities of 100% with the porcine interleukin -2 gene published in the GenBank . The eDNA of porcine interleukin 2 was cloned successfully. Then used the recombinant plasmid was cloned into eukaryotic expression vector pPICZaA and eukaryotic expression vector pPICZc^A -PoIL - 2 contained mature protein gene of porcine interleukin 2 was constructed. The recombinant expression vector was transformed into Pichia Pastoris X - 33 yeast strain by electroporation. The transformants were induced to secretive express and SDS - PAGE showed recombinant porcine interleukin 2 express system was established successfully
出处
《黑龙江畜牧兽医》
CAS
北大核心
2008年第12期13-15,共3页
Heilongjiang Animal Science And veterinary Medicine
