利用转基因技术生产C-反应蛋白的初步研究

A Study on Producing C Reactive Protein by Transgene Technology

利用转基因技术来获得C-反应蛋白,为廉价生产C-反应蛋白检测试剂盒奠定实验基础,同时初步建立一套利用转基因花生生产外源药用蛋白的实验技术体系。构建真核表达载体PBI121-CRP,利用根癌农杆菌介导法导入黑花生,使其在花生中表达,经分子生物学检测后,获得可表达CRP的转基因花生植株。其结果成功构建了真核表达载体,初步建立了黑花生再生体系,含目的基因的重组质粒转入根癌农杆菌后,共浸染105个外植体,经卡那霉素抗性筛选后得到56株抗性苗,PCR检测有13株呈阳性,经PCR-Southern杂交后,有2株出现杂交信号。初步建立黑花生再生体系,并首次成功地将CRP基因导入到植物(黑花生)中。

To obtain C-reactive protein by using transgenic technology providing a foundation for cheaply producing CRP detection regent kits,and to establish a laboratorial technology system of producing extrinsic medical protein.The eukaryotic expression vector PBI121-CRP was constructed,then transformed into peanut（Arachis hypogaea L.） Heiba920 by agrobacterium-mediated transformation for expression.After the molecular detecting,the transformed plants were obtained.The eukaryotic expression vector was successfully constructed,a highly effective regeneration system from ＂Hei＂ peanut in vitro was developed,the recombinant plasmid was antibiotic successfully transformed into agrobacterium.105 explants were dip-clyed and 56 antibiotic plants were obtained,which 13 plants were positive by PCR,only 2 plants showed cross signal by PCR-Southern-blotting.The regeneration system of ＂Hei＂ peanut was established and transgenic plants were obtained successfully for the first time.