Construction of an antisense rna expressing vector pas 1 plasmid and its inhibition on m mulv replication

Lu B.; Ji W.; Li C.; Song H.

Journal of Shanghai Medical University 19(2): 105-108


Accession: 007154735

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A retrovirus recombinant expressing antisense RNA, pAs-1 plasmid was constructed. The 1.38 kb M-MuLVpol gene fragment was isolated from pMov-3 and then ligated into pMNC vector in reverse orientation. The antisense RNA could be expressed under the CMVIE promoter. The pAs-1 plasmid was introduced into A9 cells by calcium phosphate-mediated transfection. The positive colonies were isolated by G418 and expanded to a cell line, Ao-. The results showed that the reverse transcriptase activity of cultural supernatants of A9- cells decreased by 65.8% comparing to that of nontreated A9 cells. It indicated that the antisense RNA could partially inhibit M-MuLV replication in A9- cells. The possibility of producing engineered antisense oligodeoxynucleotides is also discussed.