Molecular cloning of highly reiterated sequence AGMr (Hind I ) - 1 in Vero cell line of African green, monkey and its preliminary application
Abstract: Highly reiterated Sequence AGMr (Hind Ⅲ)-l was isolated from genome of Vero cell line derived from African green monkey and cloned into plasmid pUC18. Sequence analysis confirmed that the insert Sequence is right but 6 out of 172 bp had mutated in the 165th generation of Vero cell. Southern Blot and Dot Blot analysis of rabies vaccine prepared by Vero cell showed that residual cell DNA(RC DNA) displayed heterogeneous size from 400 ~ 5000 bp, and that the amount of RCDNA can be determined specifically and sensitively (minimun 4 pg). The fact that AGMr (Hind Ⅲ )-l Sequence represents about 20% of the genome DNA and repeats 6.8 × 106times and is rare in Primates except in members of the same genus also suggested that the recombinant plasmid could be applied as probe for detecting RC DNA existed in vaccine prepared by Vero cell and maybe used for identifying test of Vero cell.