Showing papers on "Restriction map published in 1976"
TL;DR: When a DNA molecule, enzymatically labelled with 32p at one end, is partially digested with a restriction enzyme labelled tdna fragments are obtained which form an overlapping series of molecules, all with a common labelled terminus, a restriction map can be constructed from an analysis of the size distribution of these molecules.
Abstract: When a DNA molecule, enzymatically labelled with 32p at one end, is partially digested with a restriction enzyme labelled tdna fragments are obtained which form an overlapping series of molecules, all with a common labelled terminus. ta restriction map can then be constructed from an analysis of the size distribution of these molecules. This technique has been used for the restriction site mapping of cloned histone DNA (h22) where as many as 35 cleavage sites may be accurately determined in a single experiment.
836 citations
TL;DR: It was possible to show that the synthetic globin DNA is a faithful copy of β-globin mRNA by comparing the nucleotide sequences known or predicted from the amino acid sequences of α- and β-chain rabbit hemoglobin.
416 citations
TL;DR: By techniques of recombination in vitro, a plasmid bearing the repressor gene (cI) of bacteriophage lambda fused to the promoter of the lac operon is constructed.
Abstract: By techniques of recombination in vitro, we have constructed a plasmid bearing the repressor gene (cI) of bacteriophage lambda fused to the promoter of the lac operon. Strains carrying this plasmid overproduce lambda repressor. This functional cI gene was reconstituted by joining DNA fragments bearing different parts of that gene. Flush end fusion techniques, involving no sequence overlap, were necessary for the construction; in certain cases, the abutting of the DNA molecules bearing ends generated by different restriction endonucleases creates a sequence at the junction which is recognized by one of the restriction endonucleases.
346 citations
TL;DR: The DNAs from five independent simian virus 40 (SV40) transformants of the BALB/c3T3 mouse cell line were digested with either the HpaII or the BamHI restriction endonuclease and the resulting fragments were fractionated by gel electrophoresis to show that the structure of the integrated SV40 DNA and/or its location in the host DNA are different in each line.
Abstract: The DNAs from five independent simian virus 40 (SV40) transformants of the BALB/c3T3 mouse cell line were digested with either the HpaII or the BamHI restriction endonuclease and the resulting fragments were fractionated by gel electrophoresis. The DNA fragments were denatured in situ in the gel and transferred to a membrane filter. Fragments containing viral DNA were detected by hybridization with high specific activity 32P-labeled SV40 complementary RNA (cRNA) synthesized in vitro. Each of the lines yielded a small number of fragments containing SV40 DNA and the fragments from each line were different. This observation shows that the structure of the integrated SV40 DNA and/or its location in the host DNA are different in each line.
203 citations
01 Jul 1976
TL;DR: Restriction endonucleases EcoRI and HindIII were used to analyze the structure of the plasmid genome responsible for the EcoRI restriction endonuclease and modification methylase, which appears to be identical to the ColE 1 plasmids except for a 1.95 kilobase pair segment.
Abstract: Genetic analyses of DNA restriction and modification mechanisms have been encumbered by the inability to rigorously select for mutant phenotypes associated with these systems. The application of restriction endonucleases has now proved to be a successful approach to the genetic analyses of small genomes that are recalcitrant to the more standard genetic techniques. Restriction endonucleases EcoRI and HindIII were used to analyze the structure of the plasmid genome responsible for the EcoRI restriction endonuclease and modification methylase. This plasmid in the original clinical isolate of Escherichia coli appears to be identical to the ColE 1 plasmid except for a 1.95 kilobase pair segment which contains these genes. A preliminary restriction map of this plasmid is presented.
149 citations
TL;DR: The sites of action of the restriction enzyme Bgl II on lambda DNA are mapped and this enzyme makes staggered cuts producing sticky ends and produces 5' terminals which are not substrates for polynucleotide kinase.
Abstract: The sites of action of the restriction enzyme Bgl II on lambda DNA are mapped. This enzyme recognises the sequence 5' ...AGATCT...3' and makes staggered cuts producing sticky ends. In lambda DNA, the second A in this sequence is methylated about 50% of the time by a bacterial methylase absent in E. coli dam. In contrast to Bgl II, Bgl I makes many cuts in lambda DNA and produces 5' terminals which are not substrates for polynucleotide kinase.
104 citations
TL;DR: To demonstrate the feasibility of this new method, the authors have inserted separately the synthetic lac operator DNA at the Bam I and HindIII cleavage sites of the plasmid pMB9 DNA.
97 citations
TL;DR: Two of the resulting plasmids, pCR1 and pCR11, have a single EcoRI cleavage site, but retain genetic determinants specifying resistance to colicin E1 and kanamycin, and thus may be useful as vectors for the cloning and amplification of DNA in bacteria.
Abstract: A general method has been developed for the deletion of restriction endonuclease sites in bacterial plasmid DNA. The procedure involves partial digestion of the covalently closed circular plasmid DNA with an appropriate restriction endonuclease under conditions which allow accumulation of unit-length linear DNA molecules, controlled digestion of the exposed 5′ ends with the λ 5′-exonuclease, and in vivo recircularization of the resulting linear DNA in a bacterial host cell. The method has been used for the deletion of one of the two EcoRI sites in the plasmid pML2 (colE1-Km). Two of the resulting plasmids, pCR1 and pCR11, have a single EcoRI cleavage site, but retain genetic determinants specifying resistance to colicin E1 and kanamycin, and thus may be useful as vectors for the cloning and amplification of DNA in bacteria.
88 citations
TL;DR: Results obtained suggest that DNA isolated from both HSV-1 andHSV-2 consists of molecules with four different sequence arrangements which are present in similar amounts, and some of the possible implications for genetic recombination are discussed.
Abstract: Summary
The mol. wt. and molar ratios of the Hind III and Hpa I fragments of HSV-1 DNA and the Eco RI fragments of HSV-2 DNA have been determined. Results obtained suggest that DNA isolated from both HSV-1 and HSV-2 consists of molecules with four different sequence arrangements which are present in similar amounts. Our explanation of the cleavage patterns of these four genome arrangements with the different restriction enzymes is presented.
Some of the possible implications of these four genome arrangements for genetic recombination are discussed.
80 citations
TL;DR: The sequences in lambda DNA in and around six sites cut by Hph, a restriction enzyme isolated from Haemophilus parahaemolyticus, are compared and two lines of evidence indicate that these bases constitute part or all of the Hph recognition site.
Abstract: The sequences in lambda DNA in and around six sites cut by Hph, a restriction enzyme isolated from Haemophilus parahaemolyticus, are compared. The enzyme produces a staggered cut around an AT or TA base pair, but the sequences immediately surroinding the cleavage sites bear no obvious relation to one another. Eight (in some cases nine) base pairs to one side of each cleavage site is the common sequence TCACC AGTGG. Two lines of evidence indicate that these bases constitute part or all of the Hph recognition site. First, mutations in this sequence prevent Hph cutting. Second, dimethylsulfate-mediated methylation of Gs and As in this site prevent cutting, whereas methylation of purines in the region between this sequence and the cleavage sites has no such effect. There is discernible 2-fold rotational symmetry neither in the common sequence nor around the cleavage sites.
77 citations
TL;DR: The isolation and some properties of the mitochondrial ribosomal RNA (mt-rRNA) of Drosophila melanogaster are reported, and a restriction map of the mtDNA is mapped which shows the position of the rRNA genes and of the A + T-rich region in the DNA.
TL;DR: The DNA's of the two small Bacillus subtilis phages, ∅29 and ∅15, were analyzed by use of two restriction endonucleases, EcoR1 and HpaI, and the order of the six fragments of∅15 DNA produced by EcoR2 digestion is proposed.
TL;DR: After short periods of labeling with [3H]thymidine, recently completed adenovirus DNA molecules were isolated and cleaved with restriction endonucleases, showing an asymmetry of radioactivity on the isolated strands of each restriction end onuclease piece.
Abstract: After short periods of labeling with [3H]thymidine, recently completed adenovirus DNA molecules were isolated and cleaved with restriction endonucleases. The strands (heavy and light) of most of the restriction endonuclease fragments were separated. The pattern of labeling clearly shows an asymmetry of radioactivity on the isolated strands of each restriction endonuclease piece. The data is consistent with replication proceeding in the 5' to 3' direction on each strand. Thus, there is an initiation point placed at or near each end of the molecule.
TL;DR: Electrophoretic analysis of EcoRI and HindIII restriction fragments of 2-mum supercoiled DNA of Saccharomyces cerevisiae indicated that this class of DNA is heterogeneous and probably consists of two types of molecules.
Abstract: Electrophoretic analysis of EcoRI and HindIII restriction fragments of 2-mum supercoiled DNA of Saccharomyces cerevisiae indicated that this class of DNA is heterogeneous and probably consists of two types of molecules. Integration of the 2-mum yeast DNA in E. coli plasmid pCR1 directly showed that existence of two types of molecules as each of these could be individually inserted into separate bacterial plasmids. The difference between the two types of 2-mum circles is due to an inversion of about 1.6 X 10(6) daltons. The inversion is flanked by a reversed duplicated sequence of 0.45 X 10(6) daltons. Possible implications of this structure are dicussed.
TL;DR: The mapping of the single Pst I site in ~bX174am3 RFI DNA and the determination of the sequence cleaved is described and this is the symmetrical hexanucleotide sequence.
TL;DR: The correlation of the genetic and functional map of the phage with the arrangement of fragments produced by the different enzymes has been established and complete cleavage maps were established for the enzymes EcoRI, Sal and Sma.
Abstract: The DNA of bacteriophage T5+ (molecular weight 76×106 dalton) has been dissected by various specific endonucleases. The restriction enzymes HindIII and EcoRI produce 16 and 7 fragments respectively, whereas Sal and Sma produce 4 fragments each. Complete cleavage maps were established for the enzymes EcoRI, Sal and Sma and an almost complete map for HindIII. Furthermore the location and size of the deletions St 20, St 14, b3, St 0 and b1 were determined. The correlation of the genetic and functional map of the phage with the arrangement of fragments produced by the different enzymes has been established.
TL;DR: The two forms of circular DNA which exist in equilibrium due to recombination between inverted repeat sequences were separated as stable clones, and a map of the targets for restriction endonucleases EcoRI, HindIII and HpaI was constructed.
Abstract: The 2 micron circular DNA from S. cerevisiae has been cloned on bacteriophage λ. The two forms of circular DNA which exist in equilibrium due to recombination between inverted repeat sequences were separated as stable clones, and a map of the targets for restriction endonucleases EcoRI, HindIII and HpaI was constructed. The circular DNAs isolated from a particular oligomycin resistant strain and its parent oligomycin sensitive strain were compared by restriction endonuclease analysis, and no difference was detected. The potential uses of cloned 2 micron DNA in determining the possible biological role of these plasmids are considered.
TL;DR: A new method for localizing restriction fragments by DNA-DNA hybridization and electron microscopy is described, and the map of the seven sites for the restriction endonuclease HindIII and the single site for endo R.HpaII on PM2 DNA was determined.
TL;DR: Simian virus 40 (SV40) DNA (strain 776) is cleaved by the restriction endonuclease from Arthrobacter luteus into 32 specific fragments including 20 large pieces designated Alu-A through T as well as 12 minor products named Alu m1 through m8 which were mapped on the SV40 genome by double digestion experiments.
Abstract: Simian virus 40 (SV40) DNA (strain 776) is cleaved by the restriction endonuclease from Arthrobacter luteus into 32 specific fragments including 20 large pieces designated Alu-A through T as well as 12 minor products named Alu m1 through m8. These were mapped on the SV40 genome by double digestion experiments. Alu fragments were treated with Hind enzymes and vice versa. Similar reciprocal digestions were also carried out with Hae III enzyme. In this way a detailed cleavage map of the SV40 genome could be constructed.
TL;DR: A pulse chase labelling technique using E. coli DNA polymerase I has been used to determine a number of restriction endonuclease cleavage maps of ØX 174 DNA, and the Hha I and Alu I maps have also been constructed.
Abstract: A pulse chase labelling technique using E. coli DNA polymerase I has been used to determine a number of restriction endonuclease cleavage maps of OX 174 DNA. In addition to verifying the accuracy of the method by confirming some previously published maps, the Hha I and Alu I maps have also been constructed.Images
TL;DR: In E. coli, several lines of evidence indicate that the multiple copies of the 5S, 16S and 23S rRNA genes are organised into transcription units containing one gene each plus a sequence coding either for tRNA1ILE or tRNA2GLU such that they are cotranscribed in the order 16S–4S–23S–5S RNA.
Abstract: ALTHOUGH certain segments of the primary sequence and secondary structure of 5S RNA seem to be strikingly similar in prokaryotes and eukaryotes1, the genetic organisation of the 5S RNA genes themselves exhibits remarkable variability. In E. coli, several lines of evidence indicate that the multiple copies of the 5S, 16S and 23S rRNA genes are organised into transcription units containing one gene each plus a sequence coding either for tRNA1ILE or tRNA2GLU such that they are cotranscribed in the order 16S–4S–23S–5S RNA (refs 2 and 3). In eukaryotes, the 5S RNA genes may be clustered at a single site as in humans4,5 or widely distributed on many chromosomes as in the case of Xenopus laevis, where the 5S genes are found on the telomeres of most, if not all, of the 18 chromosomes6. Among the eukaryotes, there is no example in which the 5S genes are tightly linked to the 18S and 28S rRNA genes. Although there has been a large number of papers reporting the cytogenetic localisation of the 5S RNA genes in various organisms, only in the case of Xenopus laevis are there data concerning the physical organisation of these genes7. Even in this instance, however, the structure is only known for a few 5S gene repeat lengths, and the long range order that might be imposed on an entire 5S DNA cluster within the chromosome remains obscure. Whereas long range order in certain satellite sequences has been described8,9, similar higher order organisation has not as yet been revealed for transcribed redundant genes.
TL;DR: Two different methods have been used to locate the recognition sites for the restriction endonuclease from Escherichia coli K12 on the genome of bacteriophage PM2 using purified fragments of PM2 DNA produced by digestion with Hin dIII.
TL;DR: A circular denaturation and restriction map of mitochondrial DNA from Neurospora crassa is presented and the position of all twelve fragments produced by restriction endonuclease Eco R I and the largest Hin III fragment along the previously established map of AT-rich sequences is shown.
Abstract: A circular denaturation and restriction map of mitochondrial DNA from Neurospora crassa is presented. The map shows the position of all twelve fragments produced by restriction endonuclease Eco R I and the position of the largest Hin III fragment along the previously established map of AT-rich sequences. The two wild type strains Em 5256 and 7A differ in the lengths of two Eco R I fragments. No difference was found between the mitochondrial mutant "poky" and its parent strain. The position of the DNA segment carrying the transcription unit for the two ribosomal RNA molecules has been determined by molecular hybridization.
TL;DR: A procedure which allows the resolution of complex genomes into discrete bands of specific DNA segments on agarose gels is described and possible applications are discussed.
TL;DR: In this paper, the location of mutations in all known ∅X174 genes were located on the fragment maps produced with restriction enzymes from Arthrobacter luteus (Alu I), Haemophilus influenzae Rd (Hind II), and Hahemophilus aegyptius (Hae III).
TL;DR: RNA-DNA hybridization experiments using these restriction fragments to obtain a more accurate map of the portion of the AAV genome represented in stable RNA are reported here.
Abstract: In previous work, linear duplex molecules of adeno-associated virus, type 2 (AAV2), DNA were cleaved with the restriction endonucleases R-EcoRI, R-HindII, and R-HindIII The physical order of the specific fragments obtained was deduced and oriented with respect to the DNA strand polarity and the direction of transcription Stable AAV RNA is transcribed only from 70% of the minus DNA strand We report here RNA-DNA hybridization experiments using these restriction fragments to obtain a more accurate map of the portion of the AAV genome represented in stable RNA The data obtained with several sets of restriction fragments annealed to either whole-cell RNA or poly(A)-containing RNA were internally consistent The AAV RNA annealed with a continuous region of the AAV DNA, beginning at 018 map units (18%) from the left end of the molecule and ending at 088 map units In addition, the restriction endonuclease BamHI was found to make one specific cleavage in AAV2 DNA at 022 map units, which is 004 map units (ie, 160 nucleotides) to the right (""down stream'') of the point corresponding to the 5' end of the viral mRNA
TL;DR: The nucleotide sequence at the cleavage site of the restriction endonuclease isolated from Arthrobacter luteus (Alu) has been determined and the order and sizes of these fragments have been determined to provide an Alu cleavage map of the SV40 genome.
Abstract: The nucleotide sequence at the cleavage site of the restriction endonuclease isolated from Arthrobacter luteus (Alu) has been determined. The endonuclease cleaves at the center of a palindromic tetranucleotide sequence to give even-ended duplex DNA fragments phosphorylated at the 5'-end. The endonuclease cleaves SV40 form I DNA into 32 fragments. The order and sizes of these fragments have been determined to provide an Alu cleavage map of the SV40 genome.
TL;DR: The above fragments also contain the sites susceptible to S1 endonuclease action, adding further support to the view that superhelical DNA can contain regions of localized interrupted secondary structure which may be capable of forming intrastrand hairpin structures if sequence relationships are favorable.
Abstract: Superhelical simian virus 40 (SV40) DNA I was reacted with N-cyclohexyl-N'-beta-(4-methylmorpholinium)ethylcarbodiimide (CMC), and the location of CMC sites was mapped using the Hin D restriction endonuclease. The use of 14C-labeled CMC allows a quantitative analysis of the binding to the respective Hin D restriction endonuclease fragments. The percentage of reactivity was 6.54% for fragment A, 3.87% for fragment B, and 2.74% for fragment G. No CMC radioactivity was detected in other fragments. This reactivity is in agreement with the evaluation of binding by buoyant density measurements. The above fragments also contain the sites susceptible to S1 endonuclease action. This adds further support to the view that superhelical DNA can contain regions of localized interrupted secondary structure which may be capable of forming intrastrand hairpin structures if sequence relationships are favorable. The possible structure-function relationships for this model are discussed with the emphasis on transcription.
TL;DR: To define the region of the viral genome required for T-antigen synthesis, DNA fragments were prepared by digesting SV40 DNA component I (DNA I) with Haemophilus parain fluenzae I (Hpa I) and parainfluenzae II (HPA II) restriction endonucleases and transferred into permissive monkey cells by means of microinjection technique.
TL;DR: From the restriction maps, together with data obtained by partial digestion, a physical map for the ribosomal transcription unit in yeast could be constructed.
Abstract: Yeast ribosomal DNA (rDNA) was digested with the restriction enzymes Hind III, Hind II and a mixture of Hind II and Hind III. The cleavage products were analyzed by electrophoresis on 1.5% agarose gels. Several distinct bands could be observed, which are derived from the redundant ribosomal transcription units. They are superimposed on a rather broad smear of background DNA, representing the heterogenous 'spacer' sequences. From the restriction maps, together with data obtained by partial digestion, a physical map for the ribosomal transcription unit in yeast could be constructed.