The effect of specific mutations at and around the gag-pol gene junction of Moloney murine leukaemia virus.
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.
Jones
D S
DS
Biology Division, National Cancer Center Research Institute, Tokyo, Japan.
Nemoto
F
F
Kuchino
Y
Y
Masuda
M
M
Yoshikura
H
H
Nishimura
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics*; D015683:Gene Products, gag; D051379:Mice; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000235:genetics*; D009154:Mutation; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000235:genetics; D010957:Plasmids; D012191:Retroviridae Proteins / Q000235:genetics*; D014162:Transfection; D014170:Transformation, Genetic
D004279:DNA, Viral; D015683:Gene Products, gag; D009841:Oligonucleotides; D012191:Retroviridae Proteins; D004262:DNA Restriction Enzymes