大鼠胶质纤维酸性蛋白（GFAP）互补脱氧核糖核酸（cDNA）在大肠杆菌中的克隆与表达
Cloning and expression of rat GFAP cDNA in Escherichia coli.
作者信息
Chekhonin V P, Pavlov K A, Shkoporov A N, Yefimov B A, Gurina O I, Dmitrieva T B
机构信息
Laboratory of Immunochemistry, Department of Biological Psychiatry, V. P. Serbskii Center of Social and Forensic Psychiatry, Moscow.
出版信息
Bull Exp Biol Med. 2007 Jan;143(1):68-71. doi: 10.1007/s10517-007-0019-9.
A cDNA fragment encoding GFAP was amplified by reverse transcription PCR from total mRNA isolated from primary culture of rat astrocytes and cloned for expression in Escherichia coli using pET-28a vector. High level of GFAP expression was confirmed by SDS-PAGE, while immunochemical identity was verified by immunoblotting. The constructed producer strain is a cheap source of GFAP and can be used for diagnostic purposes.
从大鼠星形胶质细胞原代培养物中分离的总mRNA通过逆转录PCR扩增编码GFAP的cDNA片段，并使用pET-28a载体克隆以在大肠杆菌中表达。通过SDS-PAGE确认了GFAP的高表达水平，同时通过免疫印迹验证了免疫化学特性。构建的生产菌株是GFAP的廉价来源，可用于诊断目的。
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