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PubMed1984

Molecular cloning and restriction mapping of a simian virus 40 deletion mutant derived from simian transformants expressing a non-karyophilic T antigen.

A replication-defective Simian virus 40 genome, with a deletion of about 120 nucleotides in the region encoding the N-terminal fourth of the large T antigen, has been isolated from the DNA of Simian cells transformed by SV40. Both the original transformants, and the murine transformants obtained by transfection with this cloned mutant DNA, produced a large T antigen displaying in immunofluorescence an exclusively cytoplasmic localization. The protein apparent molecular mass (83 kDa) was about 6% smaller than that of normal karyophilic large T. Restriction analysis showed that the deletion eliminated two close HinfI sites, at nucleotides 4459 and 4376 (map unit 0.50).

Animals↗
PubMed1983

A restriction map and analysis of the terminal redundancy in the group a streptococcal bacteriophage SP24.

The DNA isolated from the group A streptococcal bacteriophage SP24 is a linear double-stranded molecule 42.0 kb in length. The DNA has been characterized by electron microscopy, and by agarose gel electrophoresis after cleavage with the restriction endonucleases SalI, BglII, XbaI, PvuI, HindIII, and BamHI. Analysis of SalI digests indicates that two fragments are present in submolar amounts and exist as a subset of sequences present in another SalI fragment. Moreover, overlapping endonuclease fragments suggested that the physical map is circular. This was confirmed when homoduplex phage DNA revealed circular structures with single-stranded tails that were 7.7% of the circumference of the genome length molecule. Tails were observed to be separated by as much as 42% of the circular homoduplex structure. These results indicate that the phage SP24 genome is terminally redundant and circularly permuted; and the data are consistent with a model in which DNA packaging into phage heads is initiated at a specific site on concatermeric DNA and proceeds sequentially to package up to five "headfuls" of DNA per concatemer.

Bacterial Proteins↗
PubMed1992

Sheep linkage mapping: restriction fragment length polymorphism detection with heterologous cDNA probes.

A selection of cattle, human and sheep cDNA probes were screened against sheep genomic DNA, cut with 10 different restriction enzymes, to assess the usefulness of these probes for restriction fragment length polymorphism (RFLP) linkage studies in sheep. Two-thirds of the cattle cDNA probes showed moderate to strong homology with sheep DNA samples, compared with less than half of the human cDNA probes at the final washing stringency chosen for the experiments. The set of probes tested detected a useful frequency of RFLPs. Fifty-seven per cent of probes showing moderate to strong homology identified RFLPs with one or more restriction enzymes. Restriction enzymes that detected RFLPs most frequently in sheep were TaqI and MspI. The results show that sheep and cattle cDNA probes, including candidate genes for production traits, identified a high frequency of RFLPs suitable for genetic mapping in sheep.

Animals↗
PubMed1984

Tissue-specific expression of major urinary protein (MUP) genes in mice: characterization of MUP mRNAs by restriction mapping of cDNA and by in vitro translation.

The major urinary proteins (MUPs) in mice are coded for by a gene family which consists of ca. 30 members. The number of MUP genes that are expressed is not known. Previous studies have shown that MUP mRNAs are present in several secretory tissues in addition to the liver, in which they were originally identified. In this paper we show, through restriction analysis of MUP cDNAs, that distinct sets of MUP mRNAs are synthesized in each of the tissues studied and that these mRNAs are most likely coded for by different genes. As is shown, MUP mRNAs of different tissues are related to an extent that precludes the use of gene-specific probes in differentiating among them. The regions of homology also include the 3' untranslated regions of MUP mRNAs. The question of differential expression was thus investigated by searching for restriction polymorphisms in MUP mRNAs. We demonstrate that subtle differences in the sequences of even scarce mRNAs can be recognized by this particular approach. In addition, it is shown that MUP mRNAs of different tissues code for different, nonoverlapping sets of polypeptides, as determined by gel electrophoresis of in vitro-translated precursors to MUPs. The relevance of these results to models of evolution of tissue-specific regulation in a multigene family is discussed.

Animals↗
PubMed1986

Tn1721 derivatives for transposon mutagenesis, restriction mapping and nucleotide sequence analysis.

New derivatives of the tetracycline-resistance transposon Tn1721 that carry resistances to chloramphenicol, tetracycline, kanamycin and streptomycin are described. These elements are provided on various plasmid vehicles and as chromosomal insertions to extend the range of targets for Tn mutagenesis. Single EcoRI sites at the ends of these transposons proved most useful for physical mapping, for the generation of new EcoRI sites in cloning experiments, for end-labelling and for sequencing of DNA adjacent to an insertion.

Base Sequence↗
PubMed1979

Physicochemical properties and restriction maps of simian adenovirus type 38 DNA.

The sedimentation constant of simian virus type 38 (SV-38) DNA was estimated to be 31.6S. The intrinsic viscosity of DNA was on average 86.5 dl/g and the length of the molecule determined by electron microscopy was 10.6 micrometer. The average mol. wt., as determined by sedimentation and viscometry, was 21.5 x 10(6), which agreed well with the value derived from the length of the molecule (21.4 x 10(6)) and with the value of 21.2 x 10(6) determined by the relative electrophoretic mobility of the DNA fragments produced by restriction endonucleases EcoR1, SalI and BglII. The buoyant density of the DNA in caesium chloride and casesium sulphate was 1.7185 and 1.4295 g/ml respectively. The melting temperature of the DNA in 1 x SSC was 93.5 degrees C. The GC content calculated from p and Tm values was 59.3%. BglII cleaves SV-38 DNA at three sites producing four fragments with mol. wt.: A, 9.3 x 10(6), B, 5.6 x 10(6), C, 3.3 x 10(6); and D, 2.9 x 10(6). After treatment with EcoR1 and SalI, SV-38 DNA is cleaved into five and six fragments respectively, with mol. wt. for EcoR1 fragments: A, 8.2 x 10(6), B, 6.5 x 10(6), C, 4.0 x 10(6), D, 1.27 x 10(6); and E, 1.07 x 10(6), and for SalI fragments: A, 6.5 x 10(6); B, 5.4 x 10(6); C, 4.2 x 10(6), D, 2.8 x 10(6); E, 2.5 x 10(6) and F, 0.25 x 10(6). The sequence of fragments within the SV-38 DNA molecules for BglII was deduced to be BDCA, and for ECOR1--BCEAD.

Adenoviridae↗
PubMed1984

Yeast regulatory gene PPR1. I. Nucleotide sequence, restriction map and codon usage.

The PPR1 gene of Saccharomyces cerevisiae controls the transcription of two unlinked structural genes URA1 and URA3. The primary structure of this eukaryotic regulatory gene and its flanking regions has been established by the dideoxynucleotide chain termination method. Our data show an open reading frame of 2712 nucleotides, corresponding to 904 amino acid residues. The 3' untranslated messenger RNA region presents consensus yeast termination and polyadenylation sequences. The pattern of codon usage in the gene is clearly random. This result is discussed in relation to protein abundance and is compared with the codon usage in 20 yeast structural and regulatory genes and with that found for Escherichia coli genes.

Amino Acid Sequence↗
PubMedDate not supplied

Restriction maps of human adenovirus types 2, 5 and 3 for BclI, ClaI, pvul and SphI endonucleases.

The sites of cleavage by BclI, ClaI, PvuI and SphI in the DNA from adenovirus (Ad) serotypes 2, 5 and 3 have been located. Certain site coordinates were in accord with nucleotide sequences already published. A small difference in size between the PvuI-E fragments from Ad2 and Ad5 confirmed the existence of a deletion in the N-terminal moiety of Ad5 hexon gene, as previously implied by interserotypic recombinants (Boursnell and Mautner, Virology 112 (1981) 198-209) and more recently by amino acid sequencing (Von Bahr-Lindström et al., Virology 118 (1982) 352-362).

Adenoviruses, Human↗