Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Restriction Mapping”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,333 records · Page 74Linked to original sources
PubMed1990

Long range restriction analysis of the bovine casein genes.

Pulsed field gel electrophoresis (PFGE) was used to analyse the organization of the bovine alpha s1, alpha s2, beta and kappa casein genes. High molecular weight DNA was prepared from fibroblasts and lymphocytes embedded in agarose and was digested with the restriction endonucleases Clal, Sall, Smal, Xhol. The digestion products were separated by PFGE, transfered to nitrocellulose filters and hybridized to probes corresponding to the cDNAs of the four bovine casein genes. The casein genes were demonstrated to be physically linked within a region of 300 kb, represented by two adjacent Xhol fragments in fibroblasts and by a single fragment in lymphocytes. A restriction map of the casein locus was derived and the order of the genes was shown to be kappa, alpha s2, beta, alpha s1.

Animals↗
PubMedDate not supplied

[The cloning of fragments of the streptomycete plasmid pSG1912 as a part of the vector pUC19].

DNA fragments of plasmid pSG1912 isolated from Streptomyces globisporus 1912 have been cloned into vector pUC19 in the E. coli cells. Stability of inheritance of hybrid derivatives in enterobacteria has been studied. The clones where recombinant plasmids are stably inherited have been chosen. The detailed restriction maps of cloned DNA fragments of pSG1912 have been constructed.

Animals↗
PubMed1989

Cloning of xcp genes located at the 55 min region of the chromosome and involved in protein secretion in Pseudomonas aeruginosa.

Pleiotropic mutations (xcp) affecting secretion of proteins in Pseudomonas aeruginosa have been previously characterized and mapped at 0 min, 55 min and 65 min. Genomic libraries of this organism have been constructed and the genes xcp-5 and xcp-54, located at the 55 min region, were cloned using the adjacent met allele as a marker, and complementation of xcp strains. From our linkage and cloning analysis, the most probable gene order in this region appears to be pyrD... xcp-5/xcp-54/met-9011/oru-314/trpF/leu-10. Restriction mapping and transposon (Tn1725) insertion mutagenesis demonstrated that: (i) the overall size of DNA necessary for xcp expression was 9kb, (ii) the two loci are not adjacent on the chromosome, and (iii) the two loci are expressed independently. The xcp-5 gene has been subcloned on a 4kb EcoRI fragment.

Bacterial Proteins↗
PubMed1998

A polymorphism in the major immediate-early gene delineates groups among cytomegalovirus clinical isolates.

Major immediate-early gene exon 4 sequences were determined at codons 161-241 and 254-397 in 25 cytomegalovirus clinical strains and compared with those of reference strains AD169 and Towne. The nucleotide sequences at codon 161-241 segregated into three groups which could be determined by restriction mapping of a 247-nucleotide amplified target. AD169 and Towne belonged to the same group. Clustered variations and group-specific amino-acid motifs found in the deduced peptide sequence of the two immediate-early (IE) exon 4 regions raised a question is to the effects of polymorphism on IE1 function and/or immunogenicity. On the basis of restriction analysis of polymerase chain reaction (PCR) products, virus isolates were also classified into four glycoprotein B (gB) genotypes. Strain distribution in IE1 and gB genotypes showed a lack of concordance of the two grouping methods, and no preferential association was observed between the clinical context or kind of specimen and IE1 or gB groups. These data lead up to further prospective studies which could provide important information on the implication of the MIE gene region in virus pathogenesis and indicate whether linkage unbalance exists in particular clinical contexts between IE1 and gB loci.

Amino Acid Sequence↗
PubMed1997

High-resolution physical map of the X-linked retinoschisis interval in Xp22.

X-linked retinoschisis (RS) is the leading cause of macular degeneration in young males and has been mapped to Xp22 between DXS418 and DXS999. To facilitate identification of the RS gene, we have constructed a yeast artificial chromosome (YAC) contig across this region comprising 28 YACs and 32 sequence-tagged sites including seven novel end clone markers. To establish the definitive marker order, a PAC contig containing 50 clones was also constructed, and all clones were fingerprinted. The marker order is: Xpter-DXS1317-(AFM205yd12-DXS7175-DXS7992) -60N8-T7-DXS1195-DXS7993-DXS7174 -60N8-SP6-DXS418-DXS7994-DXS7995-DXS7996-+ ++HYAT2-25HA10R-HYAT1-DXS7997-DXS7998- DXS257-434E8R-3542R-DXS6762-DXS7999-DXS 6763-434E8L-DXS8000-DXS6760-DXS7176- DXS8001-DXS999-3176R-PHKA2-Xcen. A long-range restriction map was constructed, and the RS region is estimated to be 1300 kb, containing three putative CpG islands. An unstable region was identified between DXS6763 and 434E8L. These data will facilitate positional cloning of RS and other disease genes in Xp22.

Chromosome Mapping↗
PubMed1988

Genome analysis of adenovirus 4a, a causative agent of pharyngoconjunctival fever and respiratory diseases in Brazil.

An outbreak of pharyngoconjunctival fever caused by adenovirus (Ad) occurred in 1976 in Belem (northern Brazil). By restriction endonuclease analysis, it was shown that the type responsible was Ad4a, a variant of Ad4 prototype (Ad4p) and previously described as causative agent of ocular diseases. In an epidemiologic study carried out in Rio de Janeiro and Belem between 1977 and 1986, 148 Ads were isolated from children with respiratory disease. All five Ad4 strains encountered during this study were also Ad4a. This type, circulating in South America, can therefore cause both ocular and respiratory diseases. No Ad4p was isolated during this study. A percent of comigrating restriction fragments of only 50% between Ad4p and Ad4a was calculated from 13 restriction enzyme patterns. The restriction maps of Ad4a for nine endonucleases are presented and compared with those of Ad4p.

Adenoviridae↗
PubMed1994

Assignment of the true and processed genes for human glycine decarboxylase to 9p23-24 and 4q12.

Southern analysis using a human glycine decarboxylase cDNA probe and genomic DNA preparations from Chinese hamster-human hybridoma cell lines demonstrated that on the segregated human chromosomes, 4 and 9, there are glycine decarboxylase cDNA-related sequences. The finding is confirmatory of the fact that fragments of both the true and processed genes for this protein have been cloned. Since one of four HindIII fragments revealed in the present Southern analysis matched requirements for the property expected from the restriction map reported for the processed gene, this was located on chromosome 4. The three remaining signals were consequently ascribed to fragments from the true gene on chromosome 9. Fluorescence in situ hybridization using the genomic clones assigned the true and processed genes to 9p23-24 and 4q12, respectively.

Amino Acid Oxidoreductases↗
PubMed1991

Growth-phase-dependent induction of 6-phosphogluconate dehydrogenase and glucose 6-phosphate dehydrogenase in the cyanobacterium Synechococcus sp. PCC7942.

In most cyanobacteria, the only known pathway for oxidation of stored carbohydrate in the dark or under energy-limiting conditions is the hexose monophosphate shunt. To determine whether the increased use of the shunt under these conditions derives from an increase in the activity level of the respective enzymes, we measured the effect of growth phase during the growth of batch cultures of Synechococcus sp. strain PCC7942 on the specific activity of 6-phosphogluconate dehydrogenase (6PGD) and glucose 6-phosphate dehydrogenase. The specific activities were constant during the exponential growth phase of the culture, but they increased about fivefold during the transition into stationary phase. As an approach to determining the level of expression at which the growth-phase-dependent regulation of 6PGD level is exerted, we constructed operon and gene fusions between the gnd gene, which encodes 6PGD, and the Escherichia coli lacZ gene, which encodes beta-galactosidase (beta Gal). Strains harboring the fusions integrated into the cyanobacterial chromosome were prepared, and the growth-phase dependence of beta Gal level was determined. The specific activity of beta Gal in cultures of both types of fusion strains increased during the transition into stationary phase, indicating that the growth-phase-dependent regulation is on the gnd mRNA level. Characterization of the growth-phase-dependent induction of 6PGD in strains carrying differing amounts of DNA upstream from the gnd structural gene led to the localization of the promoter and the regulatory site on the restriction map of the gene, whose sequence has previously been determined.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Division↗
PubMed1991

The genetic structure of natural populations of Drosophila melanogaster. XXII. Comparative study of DNA polymorphisms in northern and southern natural populations.

Restriction map variation in four gene regions (Adh, Amy, Pu and Gpdh) was surveyed for 86 second chromosomes from northern (Aomori) and southern (Ogasawara) Japanese populations of Drosophila melanogaster (43 chromosomes from each population). The regions examined cover a total of 62 kilobases. Estimates of nucleotide diversity (pi) were approximately constant across the gene regions and populations examined. The distribution of restriction site polymorphisms was compatible with the expectation from the neutral mutation-random genetic drift hypothesis, but insertion/deletion polymorphisms were not consistent with it. While the two populations shared a majority of restriction site polymorphisms, frequencies of individual restriction site variants were significantly different between the two populations at 7 out of 35 segregating sites. In addition, an insertion in the Amy region was found in 15 chromosomes from the Ogasawara sample but absent in the Aomori sample. A considerable difference was observed in the number of rare insertions and deletions between the two populations. The numbers of aberrations uniquely represented were 16 in the Ogasawara sample and only 3 in the Aomori sample. These findings suggest that the two populations were differentiated from each other to some degree by means of random genetic drift and/or other factors.

Alcohol Dehydrogenase↗
PubMed1988

Characterization of SE-3, a virulent bacteriophage of Saccharopolyspora erythraea.

SE-3 is a virulent bacteriophage isolated from a large-scale culture of Saccharopolyspora erythraea, an erythromycin producer. The host range of the phage is narrow, limited to some strains of this species. Another strain of Sac. erythraea, and a strain of Sac. hirsuta, are able to adsorb phage particles but do not sustain their complete multiplication. SE-3 is closely related to the phage SE-5 as shown by DNA restriction mapping. The differences between SE-3 and SE-5 genomes are apparently limited to two DNA segments flanked by short inverted repeats, visualized by electron microscopy.

Bacteriophages↗
PubMed1994

Homology among Treponema denticola plasmids.

Three of 16 isolates of Treponema denticola were found to contain small (2.0-2.7 kb) cryptic plasmids. These were pTD1 from T. denticola ATCC 33520, pTD2 from strain T32A, and pTD3 from strain D3A1. These plasmids were characterized by restriction mapping and cloned into E. coli plasmid pUC19. Extensive homology between these plasmids was revealed by Southern blot, and single-stranded DNA regions were found by neutral Southern blots and S1 nuclease mapping. These plasmids were not found in serovars usually associated with human periodontal disease nor are they universal in all T. denticola strains and serovars.

Blotting, Southern↗
PubMed1989

Construction of an Escherichia coli-Clostridium perfringens shuttle vector and plasmid transformation of Clostridium perfringens.

A stable shuttle vector which replicates in Escherichia coli and Clostridium perfringens was constructed by ligating a 3.6-kilobase (kb) fragment of plasmid pBR322 with C. perfringens plasmid pHB101 (3.1 kb). The marker for this shuttle plasmid originated from the 1.3-kb chloramphenicol resistance gene of plasmid pHR106. The resulting shuttle vector, designated pAK201, is 8 kb in size and codes for resistance to 20 micrograms of chloramphenicol per ml in both E. coli and C. perfringens. Following shuttle vector construction in E. coli, plasmid pAK201 was transformed into E. coli HB101 and C. perfringens ATCC 3624A, using intact cell electroporation. The transformation frequencies were 10(6) and 10(4) transformants per microgram of DNA in E. coli and C. perfringens, respectively. Restriction enzyme analysis of the chimera isolated from transformants of both microorganisms suggested that the plasmids were identical. Reciprocal transformation experiments in E. coli and C. perfringens indicated no difference in transformation frequency. Plasmid pAK201 was stable in C. perfringens following repeated transfer in the absence of chloramphenicol pressure. The restriction map of plasmid pAK201 shows six unique cut sites which should be useful for future genetic analysis and C. perfringens gene library construction.

Chloramphenicol↗
PubMed1994

Localization of the gene (RSN) coding for restin, a marker for Reed-Sternberg cells in Hodgkin's disease, to human chromosome band 12q24.3 and YAC cloning of the locus.

A novel 160-kDa intermediate filament associated protein, named restin (Reed-Sternberg intermediate filament associated protein), is specifically expressed in the malignant cells of Hodgkin's disease and anaplastic large cell lymphoma (Ki-1 lymphoma). The combination of chromosomal R-banding and fluorescence in situ hybridization (FISH) with the use of two fluorescent dyes, fluorescein isothiocyanate and propidium iodide, allowed simultaneous detection of the hybridized DNA sequence and chromosomal R-banding. By this technique, the gene coding for restin (RSN) was assigned to chromosome region 12q24.31-->q24.33, while localization of the alpha-2-macroglobulin receptor (A2MR) was refined to 12q13.1-->q13.3. To further analyze the restin gene, a 500-kb YAC clone containing the gene was isolated and analyzed. A restriction map of this area is presented.

Chromosome Banding↗
PubMed1992

PFGE mapping and RFLP analysis of the S/D region of the mouse H-2 complex.

We have constructed a long range restriction map of the S/D segment of the mouse H-2 complex by pulsed field gel electrophoresis and hybridization with mouse cDNA probes to Bf and Tnfa genes and human cDNA probes to BAT2, BAT3, BAT4, BAT5, and BAT6 genes which have recently been mapped to the human HLA complex between C2 and HLA-B. The distance between the mouse C2 and Tnfa genes was found to be approximately 350 kilobases. The position of the mouse Bat genes in this map were found to be comparable to the position of the BAT genes in the human HLA complex. A panel of recombinant mouse strains was also examined by restriction fragment analysis with probes detecting the Hsp70, Bat5, and Tnfa genes. The results indicate that recombination in this segment is not random. No recombinants were found with crossovers between the C2 and Hsp70 genes and only one recombinant was found with a crossover between Tnfa and H-2D. In contrast, the crossover sites of 16 recombinants were mapped between the Hsp70 and Tnfa genes. Seven of these recombinants were found to have crossovers between Hsp70 and Bat5 and three recombinants were found to have crossover sites between Bat5 and Tnfa.

Animals↗
PubMed1990

Heterogeneity among the 2 microns plasmids in Saccharomyces cerevisiae: a new sequence for the REP1 gene.

Some species of yeasts contain naturally-occurring circular DNA plasmids. The most studied of these plasmids is the 2 microns circle of Saccharomyces cerevisiae. Three variants of this plasmid, Scp1, Scp2 and Scp3, have been described according to their restriction maps [Cameron et al., Nucleic Acids Res. 4 (1977) 1429-1448; Livingston, Genetics 86 (1977) 73-84]. The entire nucleotide (nt) sequence of the Scp1 variant from strain A364A has been published [Hartley and Donelson, Nature 286 (1980) 860-864]. We report here the nt sequence of the 2 microns plasmid REP1 gene from S. cerevisiae strain SKQ2n. According to the restriction analysis, this plasmid is the Scp3 variant previously described. The only observed differences between the Scp1 and Scp3 variants were the loss of one EcoRI restriction site and an apparent deletion in Scp3. The nt sequence we report differs significantly from the previously published one for Scp1. The differences correspond to 128 (about 8.5%) substituted, deleted or additional nt of 1510 nt compared. These differences affect the coding region (8%) as well as the noncoding regions (9.7%). Regarding the putative encoded proteins, 38 (about 10%) amino acids (aa) are modified or deleted in our sequence and 11 are added. Most of these aa modifications are not randomly distributed but are concentrated in certain regions. These observations are indicative of important intraspecific evolution between the two 2 microns plasmid variants considered, as well as of conservative selection pressure on some domains of the REP1 protein.

Amino Acid Sequence↗
PubMed1999

Detection of single-stranded cohesive ends in the genome of Bacillus thuringiensis temperate phage KK-88.

Cohesive ends (cos sites) were detected in the genome of temperate KK-88 phage in Bacillus thuringiensis after analysis of the phage DNA generated by the 3'-5' exonuclease activity of Klenow fragment of DNA polymerase I. In addition, unlike the 5'-protruding ends of coliphage lambda genome, the ends of KK-88 phage genome were found to be 3'-protruding. The restriction map of the phage genome was also constructed on the basis of position of the cos fragments.

Bacillus thuringiensis↗
PubMed2002

[DNA technology for diagnosis and characterization of agricultural animal pathogenic viruses].

Use of recombinant DNA for the development of diagnostic and therapeutic and preventive drugs became one of the priority trends in modern experimental veterinary. This paper discusses modern methods of virus analysis based on the DNA technologies: restriction mapping, nucleic acid hybridization, and polymerase chain reaction. Examples of utilization of these methods for clinical diagnosis and research of animal viruses are offered.

Animal Diseases↗