Showing papers on "Restriction map published in 1981"
TL;DR: A multipurpose cloning site has been introduced into the gene for beta-galactosidase on the single-stranded DNA phage M13mp2 with the use of synthetic DNA and two restriction endonuclease cleavage sites in the viral gene II were removed by single base-pair mutations.
Abstract: A multipurpose cloning site has been introduced into the gene for beta-galactosidase (beta-D-galactosidegalactohydrolase, EC 3.21.23) on the single-stranded DNA phage M13mp2 (Gronenborn, B. and Messing, J., (1978) Nature 272, 375-377) with the use of synthetic DNA. The site contributes 14 additional codons and does not affect the ability of the lac gene product to undergo intracistronic complementation. Two restriction endonuclease cleavage sites in the viral gene II were removed by single base-pair mutations. Using the new phage M13mp7, DNA fragments generated by cleavage with a variety of different restriction endonucleases can be cloned directly. The nucleotide sequences of the cloned DNAs can be determined rapidly by DNA synthesis using chain terminators and a synthetic oligonucleotide primer complementary to 15 bases preceeding the new array of restriction sites.
2,360 citations
TL;DR: Two linear deoxyribonucleic acid plasmids were isolated from the yeast Kluyveromyces lactis IFO 1267 and a single chromosomal gene was found which was responsible for the resistance to the K. lactis killer.
Abstract: Two linear deoxyribonucleic acid plasmids, designated pGK11 and pGK12, were isolated from the yeast Kluyveromyces lactis IFO 1267. pGK11 and pGK12 had molecular weights of 5.4 X 10(6) and 8.4 X 10(6), respectively. Both plasmids possessed the same density of 1.687 g/cm3, lighter than the densities of mitochondrial (1.692 g/cm3) and nuclear (1.699 g/cm3) deoxyribonucleic acids. A restriction map of pGK11 was constructed from digestions by EcoRI, HindIII, PstI, and BamHI. pGK12 was cleaved by EcoRI into seven fragments and by BamHI into two fragments K. lactis IFO 1267 killed Saccharomyces cerevisiae sensitive and killer strains and certain strains of Saccharomyces italicus, K. lactis, Kluyveromyces thermotolerans, and K. vanudenii. All K. lactis strains lacking the pGK1 plasmids were nonkillers. A hybrid was constructed between K. lactis IFO 1267 and a nonkiller K. lactis strain lacking the plasmids and subjected to tetrad analysis after sporulation. The killer character was extrachromosomally transmitted in all tetrads in association with the pGK1 plasmids. The double-stranded ribonucleic acid killer plasmid could not be detected in any K. lactis killer strains. It is thus highly probable that the killer character is mediated by the linear deoxyribonucleic acid plasmids. A single chromosomal gene was found which was responsible for the resistance to the K. lactis killer.
269 citations
TL;DR: A model of amplification in which additional rounds of replication are specifically initiated within the central gene-containing regions, followed by bidirectional replication in the absence of discrete termination sites is suggested.
232 citations
TL;DR: The DNA sequence of the filamentous phage f1, consisting of 6407 nucleotides, has been determined, showing a near identity of these two phage (there are only 59 nucleotide differences).
170 citations
143 citations
TL;DR: Cloned circular unintegrated mouse mammary tumor virus (MMTV) DNA from infected rat hepatoma cells in bacteriophage lambda was cloned and viral gene expression was increased by the addition of dexamethasone.
142 citations
TL;DR: A complete collection of fragments of Epstein-Barr virus DNA, obtained by cleavage with restriction endonuclease Eco RI, has been cloned and identified within the viral genome of each cloned DNA fragment.
Abstract: A complete collection of fragments of Epstein-Barr virus DNA, obtained by cleavage with restriction endonuclease Eco RI, has been cloned. Fourteen different internal fragments of the virus genome, derived from linear virion DNA of the B95-8 strain, and sequences corresponding to the terminal regions of virion DNA, derived from intracellular circular EBV DNA isolated from 895-8 cells, were cloned. Sizes of fragments were determined by agarose gel electrophoresis and their sum leads to an estimated molecular weight of 110 x 10(6) for virion DNA. Large Eco RI DNA fragments of special interest were also cloned in cosmids using another source of EBV DNA, that is, to circular viral DNA derived from Raji cells. In order to provide a set of overlapping sequences, all the 29 internal Bam HI fragments of B95-8 virion DNA were cloned in pBR322. The map location within the viral genome of each cloned DNA fragment was identified by hybridizing to blots of virion DNA cleaved with several different restriction endonucleases.
129 citations
TL;DR: The chromosomal beta-lactamase (penicillinase, penP) gene from Bacillus licheniformis 749/C has been cloned in Escherichia coli and it is established that B. subtilis is capable of carrying out the post-translational proteolytic cleavage(s) to convert the membrane-bound precursor enzyme into the soluble extracellular form.
Abstract: The chromosomal beta-lactamase (penicillinase, penP) gene from Bacillus licheniformis 749/C has been cloned in Escherichia coli. The locations of the target sites for various restriction enzymes on the 4.2-kilobase EcoRI fragment were determined. By matching the restriction mapping data with the potential nucleotide sequences of the penP gene deduced from known protein sequence, we established the exact position of the penP gene on the fragment. A bifunctional plasmid vector carrying the penP gene, plasmid pOG2165, was constructed which directs the synthesis of the heterologous beta-lactamase in both E. coli and Bacillus subtilis hosts. The protein synthesized in E. coli and B. subtilis is similar in size to the processed beta-lactamase made in B. licheniformis. Furthermore, the beta-lactamase made in B. subtilis is efficiently secreted by the host into the culture medium, indicating that B. subtilis is capable of carrying out the post-translational proteolytic cleavage(s) to convert the membrane-bound precursor enzyme into the soluble extracellular form.
128 citations
TL;DR: The data suggest that IgD expression in mu(+)delta(+) B cells does not involve a V(H)-to-C(delta) DNA switch rearrangement, and proposes that two alternative mechanisms may be used in the expression of IgD molecules-RNA splicing in B cells and DNA rearranged in plasma cells.
Abstract: From a library of mouse sperm DNA, we have isolated two overlapping clones which contain the Cδ gene. One of these clones also contains the Cμ gene. The Cδ gene is separated from the Cμ membrane exons by approximately 2 kilobases (kb) of DAN. The Cδ gene was identified by (a) hybridization to poly(A)+RNA prepared from the IgD-producing rat plasma cell tumor IR731, and (b) homology of a translated nucleotide sequence to the amino acid sequence of the human δ chain. The Cδ gene spans 8 kb of DNA in the germ line. Plasmid subclones of the Cδ gene were used as probes in Southern and RNA blot experiments. RNA blot analysis of cytoplasmic poly(A)+RNA from IR731 and a μ+δ+ B-cell hybridoma revealed 1.6- and 2.7-kb δ mRNA species with different 3′ ends, which presumably encode the secreted and membrane-bound forms, respectively, of the δ chain. Southern blot analysis of DNA from two μ+δ+ lymphomas revealed that the Cδ gene is in the germ-line configuration in each case. Restriction map analysis of Cμ and Cδ genomic clones isolated from a library of normal μ+δ+ B-cell DNA also gave no evidence for DNA rearrangement in the region between the Cμ and Cδ genes. Taken together, these data suggest that IgD expression in μ+δ+ B cells does not involve a VH-to-Cδ DNA switch rearrangement. We propose that simultaneous expression of Cδ and Cδ with a single VH gene is mediated by two alternative routes of RNA processing of a primary nuclear transcript which contains the VH, Cμ, and Cδ genes. In contrast, analogous experiments with myeloma IR731 DNA revealed that the Cμ gene has been deleted from the myeloma DNA and that the Cδ gene has undergone DNA rearrangement, presumably including a switch recombination of the VH gene from the Cμ to the Cδ gene. These results indicate that two alternative mechanisms may be used in the expression of IgD molecules—RNA splicing in B cells and DNA rearrangement in plasma cells.
123 citations
TL;DR: Double-stranded DNA copies of the single-Stranded genomic RNA of foot and mouth disease virus have been cloned into the Escherichia coli plasmid pBR322 and in an appropriate host the synthesis of antigenic polypeptide can be demonstrated by radioimmunoassay.
Abstract: Double-stranded DNA copies of the single-stranded genomic RNA of foot and mouth disease virus have been cloned into the Escherichia coli plasmid pBR322. A restriction map of the viral genome was established and aligned with the biochemical map of foot and mouth disease virus. The coding sequence for structural protein VP1, the major antigen of the virus, was identified and inserted into a plasmid vector where the expression of this sequence is under control of the phage lambda PL promoter. In an appropriate host the synthesis of antigenic polypeptide can be demonstrated by radioimmunoassay.
121 citations
TL;DR: The structural gene for alkaline phosphatase of Escherichia coli was cloned into the PstI site of pBR322, from a transducing bacteriophage, lambda p(phoA-proC), and the restriction map of the plasmid was established.
Abstract: The structural gene for alkaline phosphatase (phoA) of Escherichia coli was cloned into the PstI site of pBR322, from a transducing bacteriophage, lambda p(phoA-proC). The restriction map of the plasmid was established. Based upon this information, several phoA deletion plasmids as well as a smaller phoA+ plasmid were constructed. The genetic map and restriction map were correlated by recombination analysis. Cells carrying one of the phoA+ plasmids overproduce alkaline phosphatase 10-fold upon phosphate limitation. However, both regulation and processing of the enzyme were found to be normal.
TL;DR: The known effects of sequence specific methylation on restriction endonucleases are compiled.
Abstract: Sequence specific DNA methylation sometimes results in the protection of some or all of a restriction endonucleases' cleavage sites. This is usually, but not always, the result of methylation of one or both strands of DNA at the site characteristic of the corresponding "cognate" modification methylase. The known effects of sequence specific methylation on restriction endonucleases are compiled.
TL;DR: The relationship of the cloned viral DNA fragments to the XbaI physical map of the viral genome is demonstrated and even though large recombinant plasmids were isolated, most if not all of theiral DNA fragments were stable during propagation in Escherichia coli HB101.
TL;DR: The estimation of the genetic variation in a natural population when the data are obtained by the use of restriction endonucleases, using a particular DNA segment to indicate the locations and the frequencies of the recognition sequence.
Abstract: We consider the estimation of the genetic variation in a natural population when the data are obtained by the use of restriction endonucleases. Under the restriction endonuclease technique, a particular DNA segment is considered and cut wherever a recognition sequence appropriate to the endonuclease occurs. We consider data generated when a random sample of homologous DNA segments is treated in this way with one or a battery of restriction endonucleases. The numbers and sizes of the fragments that result indicate the locations and the frequencies of the recognition sequence (or, with a battery of restriction endonucleases, of each recognition sequence). These frequencies in the sample form the basis for an estimate of the amount of genetic variation in the population.
TL;DR: It is concluded that there exist at least 11 distinct genes of gene-like sequences of the interferon-alpha type in the human genome, of which most likely represents an allelic variant, and at least five pseudogenes distantly related to the interFERon- alpha genes.
TL;DR: Eleven of the endogenous proviruses of white leghorn chickens have been mapped with restriction endonucleases and specific nucleic acid hybridization reagents and are more closely related to RAV-O than to ASV, based on restriction maps and on hybridization with reagents specific for the 3′ ends of R AV-O and ASV.
TL;DR: The relative abundance of restriction site variants was highly conserved in 12 laboratory strains ofMus musculus, in embryonic and adult tissues of a single inbred strain, and in the SC-1 cell line of feral mouse origin, but appeared to differ in a feral Japanese substrain, Mus musculus molossinus.
Abstract: The structural organization of intracisternal A-particle genes has been studied, using isolates from a mouse gene library in lambda phage Charon 4A. The predominant gene form among the isolates was 7.3 kilobases (kb) in length. R-loops between the 7-kb (35S) A-particle genomic ribonucleic acid and several of these genes were colinear, with no visible evidence of intervening deoxyribonucleic acid sequences. One recombinant was found with an A-particle gene that contained a 1.7-kb deletion. Using the deletion as a reference, the deoxyribonucleic acid and ribonucleic acid homology regions were localized with respect to one another and to the restriction map: the 5' terminus of the ribonucleic acid was several hundred base pairs within the 5' end of the deoxyribonucleic acid homology region. Restriction endonuclease fragments encompassing the 5' and 3' regions of one 7.3-kb gene were separately subcloned into pBR322. Heteroduplexes between the two subclones revealed an approximately 300-base pair segment of terminally redundant sequences. The cloned 3' fragment hybridized with restriction fragments from the 5' end of several other A-particle genes, demonstrating the presence of common (though not necessarily identical) terminally repeated sequences. A-particle genes varied in the occurrence of specific restriction sites at characteristic internal loci. However, heteroduplexes between several variant 7.3-kb genes showed continuous homology regions even when spread under stringent hybridization conditions. The relative abundance of restriction site variants was highly conserved in 12 laboratory strains of Mus musculus, in embryonic and adult tissues of a single inbred strain, and in the SC-1 cell line of feral mouse origin, but appeared to differ in a feral Japanese substrain, Mus musculus molossinus. Some evidence suggests that subsets of A-particle genes may have similar flanking sequences. The results are discussed in terms of the evolution of this multigene family.
TL;DR: All but one of the PstI restriction fragments from mung bean, pea, and spinach chloroplast DNAs have been stably cloned into pBR322 and the identity of the cloned fragments to nativechloroplast DNA restriction fragments is demonstrated by restriction analysis and the ability to construct detailed restriction maps of the mungbean and pea chloropleft genomes.
TL;DR: Restriction mapping of embryonic Drosophila melanogaster DNA showed that the two genes probably occur as single copies and are closely linked, and the four heat-induced genes are most likely organized in individual transcription units.
TL;DR: Direct restriction analysis of the human genome, using the Southern transfer technique and hybridization with a human fibroblast interferon (IFN-beta) complementary DNA insert probe, revealed the presence of a single gene; no additional closely related IFN- beta genes could be detected.
Abstract: Direct restriction analysis of the human genome, using the Southern transfer technique and hybridization with a human fibroblast interferon (IFN-beta) complementary DNA insert probe, revealed the presence of a single gene; no additional closely related IFN-beta genes could be detected. A lambda-linked human gene library (Lawn et al., Cell, 15, 1157-1174 (1978)) was screened using the cDNA probe. Out of 600,000 recombinant phage examined, one single clone bearing interferon sequences was obtained. Restriction analysis of the relevant region revealed an identical restriction map as obtained for the IFN-beta 1 cDNA clones. No intervening sequences could be detected, either in the coding or the non-coding regions of the gene.
TL;DR: In this article, the authors used Colony Filter Hybridization and restriction mapping to demonstrate that a 620 NTP long insert in a plasmid clone (pSV2) represents the almost full length structural gene coding for a precursor to the seminal vesicle secretion protein IV (SVS IV).
Abstract: The abundant class of poly(A+)RNA [poly(A+)RNA11S] from rat seminal vesicle was used to synthesize ds-cDNA11S. The ds-cDNA11S was inserted and cloned into the Pst I site of pBR-322 using E. coli RR1 as host. Colony filter hybridization and restriction mapping was used to demonstrate that a 620 NTP long insert in a plasmid clone (pSV2) represents the almost full length structural gene coding for a precursor to the seminal vesicle secretion protein IV (SVS IV). The entire insert was sequenced and the coding region was matched with the known amino acid sequence. Most of the signal peptide sequence was derived from the DNA sequence. The insert in pSV2 was labelled and used to study the effect of testosterone on the accumulation of mRNA SVS IV. Administration of testosterone to castrated rats resulted in the induction of mRNA SVS IV from a few molecules per cell to levels of over 100,000 after 96 h of hormone treatment.
TL;DR: The organization of the genes coding for 18 S, 5·8 S and 26 S ribosomal RNAs in the nematode Caenorhabditis elegans is characterized and no difference is found in the major Ribosomal DNA restriction endonuclease cleavage patterns between two interbreeding strains of C. elegans.
TL;DR: Detailed restriction enzyme mapping and electron microscopic analysis showed that one of the substitution loops corresponds to an inversion of one ofthe two long terminal repeat units and adjacent cellular sequences in C60, and at least part of this region was shown to contain SSV-specific sequences not shared by SSAV.
Abstract: Closed circular viral DNA of simian sarcoma virus (SSV) and simian sarcoma-associated virus (SSAV) obtained from acutely infected dog cells was purified on preparative agarose gels, cleaved with EcoRI, and cloned in the phage lambda vector Charon 21A. The cloned 9-kilobase SSAV genome (B11) has the same restriction map as the bulk of the unintegrated linear SSAV DNA intermediate. Heteroduplex analysis between an SSV clone (lambda-C60) and an SSAV clone (lambda-B11) showed two substitution loops and one deletion loop. By using detailed restriction enzyme mapping and electron microscopic analysis, we showed that one of the substitution loops corresponds to an inversion of one of the two long terminal repeat units and adjacent cellular sequences in C60. The other substitution loop mapped close to the 3' long terminal repeat. At least part of this region was shown to contain SSV-specific sequences not shared by SSAV. The 1.9-kilobase deletion mapped at 3.5-5.5 kilobases of the linear SSAV genome, corresponding to most, if not all, of the pol gene.
TL;DR: This work has picked 12 actin recombinants from a genomic library, and at the level of restriction enzymes mapping these represent nine different genes, indicating that these recombinant were picked from a pool of at least 20 different genes.
Abstract: By three different lines of evidence there are approximately 20 copies of actin genes in the human genome. Firstly, the rate of hybridisation of a mouse actin probe to human DNA indicates that there are a minimum of 20 complementary copies of the actin sequence per genome. Secondly, this probe hybridises to 17-20 bands in Southern blots of restriction enzyme digests of total human DNA. Most of these bands hybridise with both 3' and 5' fragments of the cDNA and are therefore likely to contain the entire gene sequence. Thirdly, we have picked 12 actin recombinants from a genomic library, and at the level of restriction enzymes mapping these represent nine different genes. Probability calculations indicate that these recombinants were picked from a pool of at least 20 different genes.
TL;DR: The pea ctDNA has been found to contain one set of ribosomal RNA genes by Southern hybridization of restriction endonuclease digest, R-loop studies, and DNA-DNA heteroduplex mapping.
TL;DR: Chloramphenicol resistance plasmid pCW7 and pC221 of Staphylococcus aureus have been characterized by the construction of detailed restriction maps and by the identification of restriction sites on both plasmids which map within either the structural gene encoding CAT or its controlling elements.
TL;DR: Mouse immunoglobulin epsilon chain gene was cloned from DNA of a hybridoma producing anti-dinitrophenyl IgE, which was constructed by fusing a spleen cell of a BALB/c mouse with a variant clone of MOPC21 myeloma (IgG1 producer).
Abstract: Mouse immunoglobulin epsilon chain gene was cloned from DNA of a hybridoma producing anti-dinitrophenyl IgE, which was constructed by fusing a spleen cell of a BALB/c mouse with a variant clone of MOPC21 myeloma (IgG1 producer). Because a given active heavy chain constant region (CH) gene is linked to a heavy chain joining segment (JH) gene at its 5' side, the expressed C epsilon gene of the hybridoma was cloned from a phage library containing partial Sau3A digests of IgE hybridoma DNA by using a J gene fragment as a probe. Among 6 X 10(5) phages screened, five positive clones were obtained and three of them were identified as C epsilon gene clones by restriction mapping, Southern blot hybridization, R-loop formation, and partial nucleotide sequence determination. The determined nucleotide sequence predicted the amino acid sequence which resembles a part of the CH3 domain of human epsilon chain. The deletion profile of the C epsilon gene in various myelomas expressing different CH genes indicates that the C epsilon gene is located between the C gamma 2a and C alpha genes. The linkage (5'-epsilon-alpha-3') was directly confirmed by molecular cloning of the overlapping chromosomal segments from newborn mouse DNA.
TL;DR: A detailed restriction endonuclease cleavage map of the plasmid pKM101 has been constructed and appears to have arisen from its clinically-isolated parent by deletion of a single DNA region which codes for three of the four drug resistances carried by R46.
Abstract: A detailed restriction endonuclease cleavage map of the plasmid pKM101 has been constructed. pKM101 plasmids containing individual Tn5 insertions were used to facilitate the ordering of restriction fragments generated by enzymes cleaving pKM101 at multiple sites. By restriction enzyme analysis, pKM101 (35.4 kilobases) appears to have arisen from its clinically-isolated parent by deletion of a single DNA region which codes for three of the four drug resistances carried by R46.
TL;DR: Restriction enzyme analyses of the total DNA and the satellite I fragment show that three HpaII sites in the fragment are completely unmethylated in sperm but fully methylated in thymus DNA, which can probably account for the lower level of methylation of sperm DNA rather than hemimethylation as previously suggested.
Abstract: Genomic DNA of calf thymus contains 15 times as much 5-methylcytosine as similar sperm DNA, but the major EcoRI repeat fragment from satellite I of thymus contains ten times as much 5-methylcytosine as the corresponding fragment from sperm DNA Restriction enzyme analyses of the total DNA and the satellite I fragment show that three HpaII sites in the fragment are completely unmethylated in sperm but fully methylated in thymus DNA Under-methylation of many sites in the satellite DNAs can probably account for the lower level of methylation of sperm DNA rather than hemimethylation as previously suggested These results are also discussed in relation to maintenance and de novo (initiation-type) methylases
TL;DR: Isolation of the corresponding genomic sequences, construction of the physical map and comparing it with the restriction maps published by Barnett et al. (1) led to the conclusion that Drosophila melanogaster had isolated the genes coding for two of the three known yolk protein precursors, YP I and YP II.
Abstract: We have isolated recombinant DNA clones coding for female specific proteins from Drosophila melanogaster. By screening with 32P-(A)+RNA from male and female flies, respectively, we were able to isolate a set of 100 cDNA clones which showed a positive hybridization signal for RNA from female flies. These clones have been rescreened with RNA isolated from fat body of two day old male and female flies. We obtained four positive cDNA clones. Isolation of the corresponding genomic sequences, construction of the physical map and comparing it with the restriction maps published by Barnett et al. (1) led us to conclude that we had isolated the genes coding for two of the three known yolk protein precursors (vitellogenins), YP I and YP II. The sequence of the YP I gene was determined. It gives rise to a protein of 48 700 dalton MW which might be cleaved to a MW of 46 700 during transport. The coding sequence is interrupted by a single intron of 75 bases in length. The proposed leader sequence starts at a region homologous to six heat shock gene sequences at the site of initiation of transcription, suggesting the existence of an 11 bp cap specific consensus sequence for Drosophila melanogaster.