Study on the mammalian cell lines of stable expression strain matrix protein of rabies virus CTN-1V

Abstract

Objective: To construct CHO cell line stably expressing rabies virus matrix protein. Methods: RT-PCR was used to amplify and isolate CTV-1V gene of rabies virus. After cloning into pCDNA5.0FRT vector, the recombinant plasmid pCDNA5.0FRT-M was constructed and then transfected into CHO cells with POG44 plasmid. The positive clones were screened by hygromycin B and the stable cell lines were identified by indirect immunofluorescence and Western blot. Results:After enzyme digestion and DNA sequencing, the recombinant expression plasmid pCDNA5.0FRT-M were transfected into CHO cells, get the visible positive cell clones, scraping positive clones were expanded in culture and defined as the second generation, After 10 generations, the results were still positive.Conclusion The CHO cell line stably expressing rabies virus matrix protein was successfully established, which lays a foundation for further study of the function of the matrix protein.