pEGFP-C1-GnRH/TRS在HeLa细胞中的遗传稳定性

TL;DRAbstract

Objective To study the genetic stability of recombinant plasmid pEGFP-C1-GnRH/TRS in HeLa cells. Methods Incubate recombinant HeLa cells with pEGFP-C1-GnRH/TRS, of passages 0, 10, 20, 30 and 40, in G418-containing and G418-free media respectively, test the screened positive clones for plasmid loss rates and observe under confocal fluorescent microscope. Extract plasmids from recombinant cells of various passages for restriction analysis, and that from recombinant cells of passage 40 with the expected result for sequencing. Results The loss rates of recombinant plasmid pEGFP-C1-GnRH/TRS in HeLa cells after subculture for 40 passages were not more than 2%. The restriction maps of plasmids in HeLa cells of various passages were identical. The GnRH/TRS sequence of plasmids in HeLa cells of passage 40 was consistent with that of passage 0. The confocal fluorescent micrograph of recombinant HeLa cells of passage 40 showed no significant change as compared with that of passage 0. Conclusion R

Chat with Paper

AI Agents for this Paper