华支睾吸虫CsSCCRO基因的表达和鉴定

Expression and identification of CsSCCRO gene of Clonorchis sinensis

目的将目的基因CsSCCRO转染Hela细胞,检测转染效率和表达水平,为进一步在细胞学水平研究编码蛋白的功能提供依据。方法将目的基因CsSCCRO克隆到真核表达载体pEGFP-N1上,通过脂质体方法转入Hela细胞,RT-PCR和荧光观察来鉴定转录和表达水平。结果真核重组质粒pEGFP-N1-CsSCCRO转入Hela细胞后稳定表达荧光蛋白;RT-PCR鉴定转染的Hela细胞,扩增产物约为780 bp,与预期值相符。结论CsSCCRO基因能够转染到Hela细胞中,并且能与GFP高效转录和翻译成融合蛋白。

Objective To transfect CsSCCRO gene to Hela cell, and detect transfection efficiency, so as to provide evidence for the encoding protein function research on the cellular level. Methods Cloned CsSCCRO gene to eukaryotic vector pEGFP-N1 and transfected the pEGFP-N1-CsSCCRO to Hela cell by the method of liposome, then identified the transfection efficiency by RT-PCR. Results Recombinant plasmid pEGFP-N1-CsSCCRO was successfully transfected to Hela cell, and fluorescin were expressed stably. A 780 bp length fragment was successfully cloned and identified by RT- PCR. Conclusion CsSCCRO gene can transfect to Hela cell and can be transcribed and translated to fusion protein with GFP efficiently.