Physical mapping and complete nucleotide sequence of the denV gene of bacteriophage T4

Journal Article · Sat Dec 01 04:00:00 EST 1984 · J. Virol.; (United States)

OSTI ID:6271565

Radany, E H; Naumovski, L; Love, J D; Gutekunst, K A; Hall, D H; Friedberg, E C

Phage T4 deletion mutants that are folate analog resistant (far) and contain deletion in the region of the T4 genome near denV have been isolated previously. We showed that one of these mutants (T4farP12) expressed normal denV gene activity, whereas another mutant (T4farP13) was defective in the denV gene. The rII-distal (right) physical endpoints of these deletions defined the limits of the interval in which the rII-proximal (left) endpoint of the denV gene should be located. The deletion endpoints were identified by restriction and southern hybridization analyses of phage derivatives containing deoxycytidine instead of hydroxymethyldeoxycytidine in their DNAs. The results of these analyses localized the rII-proximal (left) end of the denV gene to a region between 62.4 and 64.3 kilobases on the T4 physical map. denV/sup +/ phage resulted from marker rescue with two of five denV/sup -/ alleles tested, using plasmids containing a 1.8-kilobase fragment from this region or a 179-base-pair terminal fragment derived from it. Sequencing of the 179-base-pair fragment from wild-type DNA showed a 130-base-pair open reading frame with its termination codon at the rII-proximal end. Confirmation that this open reading frame is part of the denV coding sequence was obtained by identifying a TAG amber codon in the homologous DNA derived from a denV amber mutant strain. This mutant strain rescued the denV/sup +/ allele from plasmids containing the wild-type sequence. An adjacent overlapping restriction fragment was also cloned, permitting determination of the remaining denV gene sequence. Based on these results, the 3' end of the coding region of the denV locus was mapped to kilobase position 64.07 on the T4 physical map, and the 5' end was mapped to position 64.48.

• Research Organization: Stanford Univ. Medical Center, CA
• OSTI ID: 6271565
• Journal Information: J. Virol.; (United States), Journal Name: J. Virol.; (United States) Vol. 52:3; ISSN JOVIA
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
BACTERIOPHAGES
CELL CONSTITUENTS
CLONING
DNA
GENE RECOMBINATION
GENES
GENETIC MAPPING
GENOME MUTATIONS
MAPPING
MICROORGANISMS
MOLECULAR STRUCTURE
MUTANTS
MUTATIONS
NUCLEIC ACIDS
NUCLEOTIDES
ORGANIC COMPOUNDS
PARASITES
PLASMIDS
VIRUSES