DNA of herpesvirus pan, a third member of the Epstein-Barr virus-Herpesvirus papio group

The DNA of herpesvirus pan, a primate B-lymphotropic herpesvirus, shares about 40% well-conserved sequence relatedness with Epstein-Barr virus (EBV) and herpesvirus papio DNAs. Labeled cloned fragments from the EBV recombinant DNA library were cross hybridized to blots of EcoRI, XbaI, and BamHI restriction endonuclease fragments of herpesvirus pan DNA to identify and map homologous sequences in the herpesvirus pan genome. Regions of colinear homology were demonstrated between 6 x 10(6) daltons and 108 x 10(6) daltons in the DNAs. The structural organization of herpesvirus pan DNA was similar to the format of Epstein-Barr virus and herpesvirus papio DNAs. The DNA consists of two domains of largely unique sequence complexity, a segment US of 9 x 10(6) daltons and a segment UL of 88 x 10(6) daltons. US and UL are separated by a variable number of tandem repetitions of a sequence IR (2 x 10(6) daltons). There was homology between DNA which mapped at 26 to 28 x 10(6) daltons and 93 to 95 x 10(6) daltons in UL. The terminal reiteration component, TR, of herpesvirus pan DNA and sequences which mapped to the left of 6 x 10(6) daltons and to the right of 108 x 10(6) daltons had no detectable homology with the corresponding regions of Epstein-Barr virus DNA.     

Colinearity between the DNAs of Epstein-Barr virus and herpesvirus papio.

Epstein Barr virus (EBV) and herpesvirus papio (HVPapio) DNAs share a common format and 40% homology. Labeled cloned fragments of EBV DNA were hybridized to blots of XbaI, EcoRI, HindIII, and SalI fragments of HVPapio DNA. EBV fragments which mapped from 2 x 10(6) to 54 x 10(6) and from 59 x 10(6) to 106 x 10(6) daltons hybridized to fragments at identical map positions in HVPapio DNA. Regions of nonhomology were demonstrated at 0 x 10(6) to 2 x 10(6), 54 x 10(6) to 59 10(6), and 106 x 10(6) to 115 x 10(6) daltons.     

DNA of Epstein-Barr virus. IV. Linkage map of restriction enzyme fragments of the B95-8 and W91 strains of Epstein-Barr Virus.

The arrangement of EcoRI, Hsu I, and Sal I restriction enzyme sites in the DNA of the B95-8 and W91 isolates of Epstein-Barr virus (EBV) has been determined from the size of the single-enzyme-cleaved fragments and from blot hybridizations that identify which fragments cut from the DNA with one enzyme contain nucleotide sequences in common with fragments cut from the DNA with a second enzyme. The DNA of the B95-8 isolate was the prototype for this study. The data indicate that (i) approximately 95 X 10(6) to 100 X 10(6) daltons of EBV (B95-8) DNA is in a consistent and unique sequence arrangement. (ii) Both termini are variable in length. One end of the molecule after Hsu I endonuclease cleavage consists of approximately 3,000 base pairs, with as many as 10 additional 500-base pair segments. The opposite end of the molecule after Sal I endonuclease cleavage consists of approximately 1,500 base pairs, with as many as 10 additional 500-base pair segments. (iii) The opposite ends of the molecule contain homologous sequences. The high degree of homology between the opposite ends of the molecule and the similarity in size of the "additional" 500-base pair segments suggests that there are identical repeating units at both ends of the DNA. The arrangement of restriction endonuclease fragments of the DNA of the W91 isolate of EBV is similar to that of the B95-8 isolate and differs from the latter in the presence of approximately 7 X 10(6) daltons of "extra" DNA at a single site. Thus, the size of almost all EcoRI, Hsu I, and Sal I fragments of EBV (W91) DNA is identical to that of fragments of EBV (B95-8) DNA. A single EcoRI fragment, C, of EBV (W91) DNA is approximately 7 X 10(6) daltons larger than the corresponding EcoRI fragment of EBV (B95-8) DNA. Digestion of EBV (W91) DNA with Hsu I or Sal I restriction endonucleases produces two fragments (Hsu I D1 and D2 or Sal I G2 and G3) which differ in total size by approximately 7 X 10(6) daltons from the fragments of EBV (B95-8) DNA. Furthermore, the EcoRI, Hsu I, and Sal I fragments of EBV (W91) and (B95-8) DNAs, which are of similar molecular weight, have homologous nucleotide sequences. Moreover, the W91 fragments contain only sequences from a single region of the B95-8 genome. Two lines of evidence indicate that the "extra" sequences present in W91 EcoRI fragment C are viral DNA and not cellular. (i) The molecular weight of the "enlarged" EcoRI C fragment of EBV (W91) DNA is identical to that of the EcoRI C fragment of another isolate of EBV (Jijoye), (ii) The HR-1 clone of Jijoye has previously been shown to contain DNA which is not present in the B95-8 strain but is present in the EcoRI C and Hsu I D2 and D1 fragments of EBV (W91) DNA (N. Raab-Traub, R. Pritchett, and E. Kieff, J. Virol. 27:388-398, 1978).     

Linkage map of Pseudomonas aeruginosa PAT.

The locations of new markers relative to markers previously mapped on the chromosome of Pseudomonas aeruginosa strain PAT were defined by generalized transduction with phage F116L and F1083. Although the marker orders of the various marker groups were deduced mainly from the results of two-factor crosses, the locations of a number of markers were confirmed by three-factor crosses. A linkage map of the chromosome of P. aeruginosa PAT was constructed which shows the relative locations of 50 genes. From the available data, the linkage maps of P. aeruginosa strains PAO and PAT appear to be similar.     

Resistance to mutagenesis of cells biochemically transformed by herpesvirus DNA fragments

LM(TK-) mouse fibroblast cells that were biochemically transformed to the dThd kinase-positive phenotype by restriction nuclease fragments of herpes simplex virus or marmoset herpesvirus DNA, all of which contained the virus dThd kinase coding region, or by HeLa S3 DNA were more resistant to mutagenesis by N-methyl-N'-nitro-N-nitrosoguanidine or 5-bromodeoxyuridine than were dThd kinase-positive LM and LM(TK-) cells. Measurements of dNTP pool sizes did not reveal relative imbalances for representative cell lines under several conditions of growth.     

Herpesvirus papio DNA is similar in organization to Epstein-Barr virus DNA.

EcoRI, HindII, SalI, nd XbaI restriction endonuclease maps of herpesvirus papio (HVPapio) DNA were derived by determining the fragment sizes and the linkage relationships between fragments generated by the different enzymes. The data indicate that HVPapio DNA has a single molecular arrangement which is similar to that of Epstein-Barr virus DNA. The size of the DNA was 110 X 10(6) to 114 X 10(6) daltons. Restriction fragments from both ends varied in the number of repeats of a 4 X 10(5)-dalton sequence, TR, and hybridized to each other. This suggests that there is an identical repeating unit, TR, at both ends of the DNA. There were usually six tandem repetitions (range, 1 to 11) of a 2 X 10(6)-dalton sequence, IR, within the DNA. IR separated the DNA into two domains of largely unique sequence complexity, a 9 X 10(6)-dalton segment, Us, and an 88 X 10(6)-dalton segment, UL. There was homology between DNA fragments which mapped at 25 X 10(6) to 29 X 10(6) to 91 X 10(6) to 95 X 10(6) daltons in UL.     

Linkage map of human chromosome 9 microsatellite polymorphisms.

Ten microsatellite markers composed of polymorphic (CA)n or (AAAT)n repeats were mapped to chromosome 9. PIC values for these markers ranged from 0.46 to 0.82. The marker at the D9S54 locus was localized to 9pter-p22 by means of a somatic cell hybrid; another marker at D9S103 was similarly localized to 9q34-qter. Two-point lod scores and individual meiotic recombination events were used to position the 10 markers relative to each other. The best order resulting from these analyses was D9S54-D9S104-[D9S52-D9S43-D9S50]-D9S53+ ++- [D9S106-D9S105]-D9S51-D9S103, with order of the loci within brackets uncertain. Two-point linkage analysis was also used to approximate the positions of the microsatellite markers relative to those of 33 markers contained in the public CEPH database (v.3) and to one other available microsatellite marker at the D9S15 locus.     

Linkage map of mouse chromosome 17: localization of 27 new DNA markers

Chromosome 17 of the laboratory variant of the house mouse (Mus musculus L.), MMU17, has been studied extensively, largely because of its involvement in the control of immune response and embryonic as well as male germ cell differentiation. A detailed linkage map of this chromosome is therefore a highly desired goal. As the first step toward achieving this goal, we have isolated, using a LINE 1 repetitive sequence as a probe, 52 anonymous DNA clones from MMU17. Twenty-seven repetitive sequence-free probes isolated from these clones displayed restriction fragment length variation among common inbred strains and could be mapped with the help of recombinant inbred strains, congenic strains, F2 segregants, or intra-t recombinants. Together with markers identified previously, the new markers can be used to construct a map of MMU17 that contains 125 DNA loci. The markers are distributed over a length of approximately 71 cM, which probably represents the entire length of MMU17. Most of the markers reside in the proximal portion of the chromosome, which contains the t and H-2 complexes; this chromosomal region is now fairly well mapped. The distal region of MMU17, on the other hand, is populated by only a few, rather imprecisely mapped markers. Molecular maps are available for most of the H-2 complex and for parts of the t complex.     

Replication of kinetoplast DNA in Trypanosoma equiperdum. Minicircle H strand fragments which map at specific locations

The mitochondrial DNA of trypanosomes, kinetoplast DNA, is a network containing thousands of topologically interlocked minicircles. Minicircles are replicated as free molecules after being detached from the network. The minicircle L strand appears to be synthesized continuously and the H strand discontinuously. This paper describes properties of Trypanosoma equiperdum minicircle H strand fragments which could be Okazaki fragments. These fragments constitute a family of molecules of discrete sizes (ranging from about 70 to 1000 nucleotides) which map to specific locations. Three of the most prominent fragments, a 73-mer, 83-mer, and 138-mer, map at contiguous or overlapping sites. Based on their position relative to the initiation site for L strand synthesis, the 73-mer may be the first Okazaki fragment to be synthesized and either the 83-mer or the 138-mer may be the second. The 5' end of the 73-mer lies within a sequence, GGGCGT, found at a similar location in minicircles of all trypanosomatid species. During the maturation of free minicircles and after their reattachment to the networks there appears to be continued extension and ligation of the H strand fragments. However, the ligation of the 73-mer, 83-mer, and 138-mer to the rest of the H strand is delayed; their eventual ligation results in covalent closure of the minicircles.     

Replication of herpesvirus DNA. III. Rate of DNA elongation.

The rate of pseudorabies virus DNA elongation was measured by three different techniques: density shift experiments, radioautography examined by light microscopy, and radioautography examined by electron microscopy. The rate of the fork movement at 37 degrees C was estimated to be approximately 1 micron/min.