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中文摘要：
为了建立稳定表达猪FcγRⅢ（Porcine Fc gamma receptorⅢ,poFcγRⅢ）的Marc-145细胞系,从PAM细胞中提取总RNA,应用RT-PCR技术获得猪FcγRⅢ和γ链的cDNA,并构建PIREShyg3-γ和pcDNA3.1-FcγRⅢ真核表达质粒;用脂质体共转染Marc-145细胞,经潮霉素B（300 mg/L）和G418（400 mg/L）共筛选获得稳定表达猪FcγRⅢ的细胞系;运用RT-PCR、玫瑰花环试验和流式细胞术对细胞系进行了鉴定。结果表明,成功构建了猪FcγRⅢ和γ链真核表达载体,建立了稳定表达猪FcγRⅢ的细胞系,表达于转染细胞表面的猪FcγRⅢ受体分子能与猪IgG特异结合。
英文摘要：
The objective of the present study is to establish the Marc-145 cell line that stably expresses porcine poFcγRⅢ（porcine Fc gamma receptor Ⅲ,poFcγRⅢ）.The total RNA was extracted from porcine alveolar macrophage（PAM）,and poFcγRⅢ and γ-chain cDNAs were cloned by RT-PCR,then they were inserted into the eukaryotic expression vectors pcDNA3.1（＋）and PIREShyg3 respectively.The marc-145 cell line was stably transfected with pcDNA3.1-poFcγRⅢ and PIREShyg3-γ plasmids by Lipofectamine2000,then the co-transfected cells were selected by HygromycinB（300 mg/L）and G418（400 mg/L）.The expression of poFcγRⅢ on transfected cells was verified through RT-PCR,rosetting test and FCM.The eukaryotic expression vectors were successfully constructed and the Marc-145 cell line with stable poFcγRⅢ expression was obtained.The Marc-145 cell transfected with the poFcγRⅢ cDNA were able to bind porcine IgG.
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