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Nucleic Acids Research, 1989, Vol. 17, No. 15 5933-5945
© 1989

MOLECULAR BIOLOGY

The effect of specific mutations at and around the gag-pol gene junction of Moloney murine leukaemia virus

David S. Jones*,+, Fumiko Nemoto, Yoshiyuki Kuchino, Michiaki Masuda1, Hiroshi Yoshikura1 and Susumu Nishimura

Biology and Biophysics Divisions, National Cancer Center Research Institute 5-1-1 Tsukiji, Chuo-ku, Tokyo 104 1Department of Bacteriology, Faculty of Medicine, University of Tokyo 7-3-1 Hongo, Bunkyo-ku, Tokyo 113, Japan

*To whom correspondence should be addressed at: Department of Biochemistry, University of Liverpool, PO Box 147, Liverpool L69 3BX, UK

Received May 26, 1989. Revised July 18, 1989. Accepted July 18, 1989.

By carrying out oligonucleotide-directed mutagenesis, in vitro, on a 3.3 kb XhoI-HindIII fragment from Moloney murine leukaemia virus Mo-MuLV proviral DNA, inserted into the phagemid pTZ19R, nine separate fragments have been prepared in which mutations have been inserted at and around the gag-pol gene junction. Using these mutant fragments Mo-MuLV proviral DNA has been reassembled and cloned into pBR322. Examination of the mutant proviral DNAs in mouse culture cells indicates that a terminator codon at the gag-pol junction is essential for function, but any of the three chain terminator codons gives an active virus. Also the region of secondary structure surrounding the terminator codon must be preserved.

+On leave of absence from Department of Biochemistry, University of Liverpool, PO Box 147, Liverpool L69 3BX, UK


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