摘要
目的 克隆并表达人心肌肌钙蛋白I (cTnI)cDNA ,为高效获得大量蛋白、制备抗体打下基础。方法 采用逆转录 聚合酶链反应 (RT PCR)和分子生物学技术克隆人cTnI基因 ,重组构建pcDNA3 cTnI真核表达载体 ,转染人胚肾(HEK) 2 93细胞 ,并用单克隆抗体检测特异性cTnI蛋白表达。结果 克隆了cTnI的全长基因 ,构建了pcDNA3 cTnI真核表达载体并成功地在真核细胞中获得了蛋白表达。结论 所表达的蛋白证明具有正确的cTnI免疫原性 。
Objective To clone and express human cardiac troponin I cDNA,and to further investigate the method of obtaining the human cardiac troponin I (cTnI) at a high level.Methods Reverse transcription polymerase chain reaction based cloning method,cell transfection,and Western blot were used.Results The human cTnI encoding cDNA was successfully cloned and inserted into pcDNA3 eucaryotic expression vector.The expressed HA cTnI fusion protein was antigenic against anti HA and anti cTnI monoclonal antibodies.Conclusion The method provides a way for the eventual goal of obtaining a source of recombinant cTnI for diagnostic use.
出处
《中华老年心脑血管病杂志》
CAS
2000年第3期170-172,共3页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
基金
国家自然科学基金!资助项目 ( 3 960 0 0 63 )
