猪IgG Ⅱ类Fc受体基因真核表达重组质粒的构建及表达  

Construction and Expression of the Eukaryotic Expression Plasmid of Porcine Immunoglobulin G Fc Receptor Ⅱ

作　　者：张玉杨[1] 张改平[2] 乔松林[2] 郭成留[1] 

机构地区：[1]河南省农业科学院畜牧兽医研究所,河南郑州450002 [2]河南省动物免疫学重点实验室,河南郑州450002

出　　处：《华北农学报》2007年第4期52-55,共4页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金(30400323)

摘　　要：利用本研究小组克隆的猪IgG Ⅱ类Fc受体cDNA(swFcγRⅡ)基因序列(DQ026064)设计引物,应用RT-PCR技术,从猪外周血白细胞cDNA中扩增了完整的960 bpswFcγRIIORF序列,利用基因重组技术将swFcγRIIORF基因亚克隆到真核表达载体pcDNA3,经PCR及双酶切鉴定:扩增出901 bp的PCR产物;KpnI/EcoR I双酶切,切出5 376和901bp DNA片段,表明重组质粒pcDNA/swFcγRⅡ构建成功。然后脂质体法转染COS-7细胞,用玫瑰花环试验鉴定了swFcγRII配体亲和特性。The Fc receptor cDNA sequence of porcine IgG Ⅱ cloned by us was used to design primers. Using RT- PCR technology, a 960 bp of swFcγRⅡ sequence covering the whole coding region was amplified from the peripheral blood leucocyte cDNA, and the was sub-cloned into the expression vector pcDNA3. Through double digestion by Kpn I/EcoR I and PCR identification, a 5 375 bp and a 901 bp of DNA fragments were obtained, indicating that the recombinant plasmid pcDNA/swFcγRⅡ was successfully constructed. The 901 bp fragment was then transfected into COS-7 cells by the lipofection method, and the ligand affinity of swFcγRⅡ was identified by rosetting procedures.

关 键 词：猪 IGG Ⅱ类Fc受体 表达载体 玫瑰花环试验 

分 类 号：S828.03[农业科学—畜牧学]