Showing papers on "Gene published in 1979"
TL;DR: This co-transformation system should allow the introduction and stable integration of virtually any defined gene into cultured cells and is likely to represent a subpopulation of competent cells which are likely to integrate other unlinked genes at frequencies higher than the general population.
1,281 citations
Book•
31 Mar 1979
TL;DR: Regulation of Plant Intercellular Communication Via Plasmodesmata and Paired-End Genomic Signature Tags are described as a Method for the Functional Analysis of Genomes and Epigenomes.
Abstract: Regulation of Plant Intercellular Communication Via Plasmodesmata.- Root-Knot and Cyst Nematode Parasitism Genes: The Molecular Basis of Plant Parasitism.- Mutagenesis of Human p53 Tumor Suppressor Gene Sequences in Embryonic Fibroblasts of Genetically-Enginered Mice.- Salicylic Acid-, Jasmonic Acid- and Ethylenemediated Regulation of Plant Defense Signaling.- Proximity Ligation: A Specific and Versatile Tool for the Proteomic Era.- Protein Overexpression in Mammalian Cell Lines.- A High-Throughput Approach To Protein Structure Analysis.- New Mass-Spectrometry-Based Strategies For Lipids.- Paired-End Genomic Signature Tags: A Method for the Functional Analysis of Genomes and Epigenomes.
959 citations
TL;DR: The complete nucleotide sequence of hepatitis B virus genome (subtype ayw) cloned in Escherichia coli has been determined using the Maxam and Gilbert method and the dideoxynucleotide method.
Abstract: The complete nucleotide sequence of hepatitis B virus genome (subtype ayw) cloned in Escherichia coli has been determined using the Maxam and Gilbert method and the dideoxynucleotide method. This sequence is 3,182 nucleotides long. Location of the nonsense codons shows that the coding capacity of the L chain is larger than the coding capacity of the Schain. Eight open regions, able to code for polypeptide chains larger than 100 amino acids, have been located. Region 6, which is the largest, covers more than 80% of the genome. The gene S which codes for polypeptide I of the Hbs Ag and was previously located between coordinates 95.1 and 73.6 is contained in region 7.
940 citations
TL;DR: Synthetic genes for human insulin A and B chains were cloned separately in plasmid pBR322 and fused to an Escherichia coli beta-galactosidase gene to provide efficient transcription and translation and a stable precursor protein.
Abstract: Synthetic genes for human insulin A and B chains were cloned separately in plasmid pBR322. The cloned synthetic genes were then fused to an Escherichia coli beta-galactosidase gene to provide efficient transcription and translation and a stable precursor protein. The insulin peptides were cleaved from beta-galactosidase, detected by radioimmunoassay, and purified. Complete purification of the A chain and partial purification of the B chain were achieved. These products were mixed, reduced, and reoxidized. The presence of insulin was detected by radioimmunoassay.
789 citations
TL;DR: This hypothetical structure and gel-blotting analysis of total embryo and myeloma DNA suggest that the somatic recombination may be accompanied by excision of an entire DNA segment between a V gene and a J DNA segment, which may generate antibody diversity by modulation of the precise recombination sites.
Abstract: The entire nucleotide sequence of a 1.7-kilobase embryonic DNA fragment containing five joining (J) DNA segments for mouse immunoglobulin kappa chain gene has been determined. Each J DNA segment can encode amino acid residues 96--108. Comparison of one of the five J DNA sequences with those of an embryonic variable (V) gene and a complete kappa chain gene permitted localisation of a precise recombination site. The 5'-flanking regions of J DNA segments could form an inverted stem structure with the 3'-non-coding region of embryonic V genes. This hypothetical structure and gel-blotting analysis of total embryo and myeloma DNA suggest that the somatic recombination may be accompanied by excision of an entire DNA segment between a V gene and a J DNA segment. Antibody diversity may in part be generated by modulation of the precise recombination sites.
730 citations
TL;DR: An adenovirus type 5(Ad5) early gene function located in early region 1 which is required for the production of early cytoplasmic mRNAs corresponding to early regions 2, 3, and 4 is identified.
Abstract: We have identified an adenovirus type 5(Ad5) early gene function located in early region 1 which is required for the production of early cytoplasmic mRNAs corresponding to early regions 2, 3, and 4. Mutant dl312 (lacks the segment between 1.5 and 4.5 map units) grows as well as wild-type virus in 293 cells (Ad5-transformed human embryonic kidney cells), but its growth is severely restricted in HeLa cells. We detect no viral RNAs in the cytoplasm of dl312-infected HeLa cells. Viral RNA sequences are present, however, in dl312-infected HeLa cell nuclei.
685 citations
TL;DR: This ‘hybrid’ gene was expressed in Escherichia coli under the control of the lac promoter and a polypeptide was produced having the size and immunological properties characteristic of mature human growth hormone.
Abstract: DNA coding for human growth hormone was constructed by using chemically synthesised DNA in conjunction with enzymatically prepared cDNA. This ‘hybrid’ gene was expressed in Escherichia coli under the control of the lac promoter. A polypeptide was produced having the size and immunological properties characteristic of mature human growth hormone.
674 citations
TL;DR: It is demonstrated that the expression of many early adenovirus mRNAs is dependent upon the activity of a pre-early viral product, which is defective in adanovirus 5 host range (Ad hr) group I mutants.
642 citations
TL;DR: The defective metabolism of sparteine was found to have a genetic basis and appears to be determined by two allelic genes at a single locus where nonmetabolisers are homozygous for an autosomal recessive gene.
Abstract: Sparteine, an antiarrhythmic and oxytocic drug, is metabolised by N1-oxidation. The sparteine-N1-oxide rearranges with loss of water to 2- and 5-dehydrosparteine. 18 (i.e., 5%) out of 360 subjects were unable to metabolise the drug. These persons, who were designated as nonmetabolisers, excreted almost 100% of the administered dose in urine as unchanged drug. The defective metabolism of sparteine was found to have a genetic basis. Sparteine-N1-oxidation appears to be determined by two allelic genes at a single locus where nonmetabolisers are homozygous for an autosomal recessive gene.
615 citations
TL;DR: Comparison of the human mitochrondrial DNA sequence of the cytochrome oxidase subunit II gene and the sequence of a corresponding beef heart protein shows that UGA is used as a tryptophan codon and not as a termination codon, and suggests that AU A may be a methionine and not an isoleucine codon.
Abstract: Comparison of the human mitochrondrial DNA sequence of the cytochrome oxidase subunit II gene and the sequence of the corresponding beef heart protein shows that UGA is used as a tryptophan codon and not as a termination codon and suggests that AU A may be a methionine and not an isoleucine codon. The cytochrome oxidase II gene is contiguous at its 5′ end with a tRNAAsp gene and there are only 25 bases at its 3′ end before a tRNALys gene. These tRNAs are different from all other known tRNA sequences.
497 citations
TL;DR: These experiments prove that in five mouse and rat cell lines the chemically induced phenotype is encoded in DNA, and the sequences specifying the transformed phenotype behave as a dominant allele in the NIH3T3 recipient cells.
Abstract: DNA was prepared from 15 different mouse and rat cell lines transformed by chemical carcinogens in vitro and in vivo. These DNAs were applied to NIH3T3 mouse fibroblast cultures by using the calcium phosphate transfection technique. DNAs of five donor lines were able to induce foci on the recipient monolayers. Ten other donor DNAs yielded few or no foci. DNAs from control, nontransformed parental cell lines induced few or no foci. Chromosomes were transfected from one donor whose naked DNA was unable to induce foci, and morphologic transformation of recipients was observed. These experiments prove that in five of these cell lines the chemically induced phenotype is encoded in DNA, and the sequences specifying the transformed phenotype behave as a dominant allele in the NIH3T3 recipient cells. The sequences encoding the transformation are likely found on a single fragment of DNA.
TL;DR: Calculations based on the divergence of the two preproinsulin nucleotide and amino acid sequences indicate that these genes are the products of a recent duplication.
TL;DR: A large number of motility mutants of the gliding bacterium Myxococcus xanthus have been isolated and analyzed by transduction, and almost all nonmotile mutants are found to be double mutants.
Abstract: A large number of motility mutants of the gliding bacterium Myxococcus xanthus have been isolated and analyzed by transduction Almost all nonmotile mutants are found to be double mutants This is explained by the existence of two parallel and almost independent multi-gene systems controlling motility, in which case at least one mutation in each system would be required eliminate motility Only one locus, called mgl, has been found to be shared by both systems Wild type cells move singly and in groups Single cells move if they carry a complete gene system A, the genes of which are described in the preceding paper Groups of cells can move if they carry a complete gene system S, but single A−S+ cells do not move Linkage analysis reveals at least 9 different loci in system S One class of S− mutants, those mutated in the locus tgl, are conditional mutants which, after contact with tgl
+ cells, become temporarily motile as cell groups Most system A mutations have little effect on fruiting but many system S mutations block it, suggesting that system S plays a role in the fruiting process
TL;DR: The data reported predict the previously unknown sequence of the signal peptide of human growth hormone and strengthens the hypothesis that these genes evolved by gene duplication from a common ancestral sequence.
Abstract: The nucleotide sequence of a DNA complementary to human growth hormone messenger RNA was cloned; it contains 29 nucleotides in its 5' untranslated region, the 651 nucleotides coding for the prehormone, and the entire 3' untranslated region (108 nucleotides). The data reported predict the previously unknown sequence of the signal peptide of human growth hormone and, by comparison with the previously determined sequences of rat growth hormone and human chorionic somatomammotropin, strengthens the hypothesis that these genes evolved by gene duplication from a common ancestral sequence. The human growth hormone gene sequences have been linked in phase to a fragment of the trp D gene of Escherichia coli in a plasmid vehicle, and a fusion protein is synthesized at high level (approximately 3 percent of bacterial protein) under the control of the regulatory region of the trp operon. This fusion protein (70 percent of whose amino acids are coded for by the human growth hormone gene) reacts specifically with antibodies to human growth hormone and is stable in E. coli.
TL;DR: The fine structure map of the yeast URA 3 gene was established by meiotic recombination, and amber nonsense mutations were located at different points on the map, and nonsense mutations reduce the messenger level without lowering its instantaneous rate of synthesis.
Abstract: The fine structure map of the yeast URA 3 gene was established by meiotic recombination, and amber nonsense mutations were located at different points on the map. The effect of the length of the labeling time on the specific radioactivity of ura 3 messenger RNA and on its repartition between poly(A)-RNA and RNA not containing poly(A) has been followed in nonsense mutants. Nonsense mutations reduce the messenger level without lowering its instantaneous rate of synthesis. The strength of the reduction depends on the position of the nonsense codon within the locus and concerns essentially the accumulation of polyadenylylated ura 3 mRNA.
Patent•
04 Jan 1979TL;DR: In this article, a method and compositions for replication and expression of exogenous genes in microorganisms are provided for the production of nucleic acids and proteins, such as commercially useful enzymes, which may have direct usefulness, or may find expression in the synthesis of drugs.
Abstract: Method and compositions are provided for replication and expression of exogenous genes in microorganisms. Plasmids or virus DNA are cleaved to provide linear DNA having ligatable termini to which is inserted a gene having complementary termini, to provide a biologically functional replicon with a desired phenotypical property. The replicon is inserted into a microorganism cell by transformation. Isolation of the transformants provides cells for replication and expression of the DNA molecules present in the modified plasmid. The method provides a convenient and efficient way to introduce genetic capability into microorganisms for the production of nucleic acids and proteins, such as medically or commercially useful enzymes, which may have direct usefulness, or may find expression in the production of drugs, such as hormones, antibiotics, or the like, fixation of nitrogen, fermentation, utilization of specific feedstocks, or the like.
TL;DR: The results indicate that DLVs represent recombinants between a common ‘vector’ related to a chicken endogenous virus and one of three types of cellular gene possibly involved in haematopoietic differentiation.
Abstract: The RNAs of seven replication-defective leukaemia virus (DLV) strains contain three types of unique sequences, which correlate with the capacity of a given virus strain to transform erythroblasts, macrophage-like cells and myeloblasts, respectively. These sequences, termed erb, mac and myb, have their counterparts in the normal DNA of avian and mammalian species. Our results indicate that DLVs represent recombinants between a common 'vector' related to a chicken endogenous virus and one of three types of cellular gene possibly involved in haematopoietic differentiation.
TL;DR: Three types of variant restriction enzyme patterns of globin DNA fragments were detected in otherwise normal individuals and can be used to derive an approximate estimate of the total number of different DNA sequence variants in man.
TL;DR: By altering the frame of recombination, this work can account for additional light chain amino acid sequences, suggesting that the V/J joining event might generate antibody diversity somatically both by using different combinations of variable and J region genes and by using alternative joining frames.
Abstract: Immunoglobulin kappa chain gene formation involves site-specific somatic recombination between one of several hundred germ-line variable region genes and a joining site (or "J segment") encoded close to the constant region gene. We have cloned and determined the nucleotide sequence of major portions of the recombination region of the mouse kappa gene and discovered a series of five such J segments spread out along a segment of DNA 2.4 kilobases from the kappa constant region gene. These J segments encode the 13 COOH-terminal amino acids of the variable region, probably including amino acids involved in the antigen combining site and in heavy/light chain contacts. The J segments also display striking sequence homology to one another in both their coding and immediately flanking sequences. Major elements of a short palindrome--CAC(TA)GTG--are preserved adjacent to the recombination sites of both variable and J region genes and constitute inverted repeats at both ends of the sequences to be joined. These palindromes can be written as a hypothetical stem structure that draws variable and J regions together, providing a possible molecular basis for the DNA joining event. Four of the J segments that we have discovered encode amino acid sequences already found in myeloma proteins. By altering the frame of recombination, we can account for additional light chain amino acid sequences, suggesting that the V/J joining event might generate antibody diversity somatically both by using different combinations of variable and J region genes and by using alternative joining frames.
TL;DR: Rabbit β-globin complementary DNA has been inserted into SV40DNA in place of the gene coding for the virus' major capsid protein, VP1, and the recombinant genome, SVGT5-RaβG, multiplies efficiently in CV1 monkey kidney cell cultures and is transcribed to yield cytoplasmic, polyadenylated hybrid mRNAs containing the β- globin coding sequence.
Abstract: Rabbit beta-globin complementary DNA (cDNA) has been inserted into SV40 DNA in place of the gene coding for the virus' major capsid protein, VP1. The recombinant genome, SVGT5-RabetaG, multiplies efficiently in CV1 monkey kidney cell cultures and is transcribed to yield cytoplasmic, polyadenylated hybrid mRNAs containing the beta-globin coding sequence. Cells propagating SVGT5-RabetaG produce substantial quantities of rabbit beta-globin polypeptide.
TL;DR: A clone of Vero cells was isolated and shown to be totally unable to synthesize interferon and insensitive to the toxic effect of poly(rI).poly(rC) treatment, and it is proposed that in Vero Cells, the gene forinterferon synthesis is defective or absent.
Abstract: Summary
A clone of Vero cells was isolated and shown to be totally unable to synthesize interferon and insensitive to the toxic effect of poly(rI).poly(rC) treatment. Cells of this clone and mouse L cells were fused by treatment with polyethylene glycol or Sendai virus. Hybrid cell clones were isolated following selection in medium containing hypoxanthine, thymidine and ouabain. The hybrids were sensitive to the antiviral effect of poly(rI).poly(rC) and synthesized mouse, but not primate, interferon. It is proposed that in Vero cells, the gene for interferon synthesis is defective or absent.
TL;DR: DNA hybridization evidence with appropriate labeled probes indicates clearly that in some instances transformation involves the integration of bacterial plasmid sequences together with the qa-2(+) gene into the N. crassa genome.
Abstract: An efficient transformation system has been developed for Neurospora crassa that uses spheroplasts and pVK88 plasmid DNA. pVK88 is a recombinant Escherichia coli plasmid carrying the N. crassa qa-2+ gene which encodes catabolic dehydroquinase (3-dehydroquinate hydro-lyase, EC 4.2.1.10) and is part of the qa gene cluster. The recipient strain carries a stable qa-2- mutation and an arom-9- mutation, thus lacking both catabolic and biosynthetic dehydroquinase activities. Transformants were selected as colonies able to grow in the absence of an aromatic amino acid supplement. These colonies were qa-2+ and had normal levels of catabolic dehydroquinase. DNA·DNA hybridization evidence with appropriate labeled probes indicates clearly that in some instances transformation involves the integration of bacterial plasmid sequences together with the qa-2+ gene into the N. crassa genome. On the basis of genetic, enzyme assay, and DNA hybridization data, at least three types of transformation events can be distinguished: (i) replacement of the qa-2- gene by the qa-2+ gene without any effect on the expression of the other genes in the qa cluster, (ii) linked insertion of a normal qa-2+ gene accompanied by inactivation of the adjacent qa-4+ gene, and (iii) insertion of a normal qa-2+ gene at an unlinked site in the N. crassa genome. This newly integrated qa-2+ genetic material is inherited in a typical Mendelian fashion. A low level of transformation has also been obtained by using linear total N. crassa DNA. Two such qa-2+ transformants are unlinked to the qa-2- gene of the recipient.
TL;DR: These BKV DNA sequences are compared with analogous regions in the SV40 and Py virus genomes in an attempt to localize viral functions for lytic growth and transformation.
TL;DR: In this article, the authors used antisera directed against a single viral gene (src), which encodes a phosphoprotein (pp60src) with the enzymatic activity of a protein kinase.
Abstract: Neoplastic transformation of cell by avian sarcoma virus is mediated by a single viral gene (src), which encodes a phosphoprotein (pp60src) with the enzymatic activity of a protein kinase. The DNAs of vertebrate species contain a highly conserved homologue of src that is also represented in the polysomal RNA of uninfected cells and, hence, may specify a normal cellular protein. We have used antisera directed against pp60src to isolate a closely related phosphoprotein (denoted vertebrate pp60) from uninfected chicken, quail, rat, and human cells. Our data indicate that vertebrate pp60 is a homologue of pp60src, highly conserved both antigenically and chemically. Moreover, the cellular protein may possess protein kinase activity similar to that associated with pp60src. We conclude that the product of src is a slightly modified analogue of a normal cellular protein.
TL;DR: It is reported that in mouse S-180 and L5178Y cell lines unstably amplified dihydrofolate reductase DNA sequences are associated with small, paired chromosomal elements denoted "double minute chromosomes," whereas in stably amplified cells of the same origin, the genes are related with large chromosomes.
Abstract: Selection of mammalian cells in progressively increasing concentrations of methotrexate results in selective amplification of DNA sequences coding for dihydrofolate reductase (tetrahydrofolate dehydrogenase, 5,6,7,8-tetrahydrofolate:NADP+ oxidoreductase, EC 1.5.1.3). In some cell variants the amplified genes are stable with growth in the absence of methotrexate, whereas in other variants the amplified genes are lost from the population. We have previously reported that in a stably amplified variant of Chinese hamster ovary cells, the genes are localized to a single chromosome. Herein we report that in mouse S-180 and L5178Y cell lines unstably amplified dihydrofolate reductase DNA sequences are associated with small, paired chromosomal elements denoted "double minute chromosomes," whereas in stably amplified cells of the same origin, the genes are associated with large chromosomes.
TL;DR: The rat serum albumin gene has been isolated from a recombinant library containing the entire rat genome cloned in the lambda phage Charon 4A and preliminary R-loop and restriction analysis has revealed that this gene is split into at least 14 fragments by 13 intervening sequences.
Abstract: The rat serum albumin gene has been isolated from a recombinant library containing the entire rat genome cloned in the lambda phage Charon 4A. Preliminary R-loop and restriction analysis has revealed that this gene is split into at least 14 fragments (exons) by 13 intervening sequences (introns), and that it occupies a minimum of 14.5 kilobases of genomic DNA.
TL;DR: The complete sequence of the iso-1-cytochrome c gene of yeast has been determined and supports a model in which translation starts at the first AUG down-stream from the 5′ terminus of the mRNA, with no other sequence requirements.
TL;DR: The amino acid sequence deduced from the DNA indicates that the surface antigen is a protein consisting of 226 amino acids and with a molecular weight of 25,398, and the portion of the gene coding for this protein apparently contains no intervening sequences.
Abstract: DNA extracted from hepatitis B virus Dane particles has been cloned in bacteria using a plasmid vector. A full-length clone has been examined by restriction endonuclease analysis, and the nucleotide sequence of an 892-base pair fragment from cloned hepatitis B viral DNA encoding the surface antigen gene is reported. The amino acid sequence deduced from the DNA indicates that the surface antigen is a protein consisting of 226 amino acids and with a molecular weight of 25,398. The portion of the gene coding for this protein apparently contains no intervening sequences.
TL;DR: A clone which contains the complete chicken ovalbumin gene, including its leader coding sequences, has been isolated and it is concluded that the minimal size of the transcriptional unit for Ovalbumin is 7.7 kilobases.
Abstract: A clone which contains the complete chicken ovalbumin gene, including its leader coding sequences, has been isolated. From electron microscopic analysis of this DNA we conclude that the minimal size of the transcriptional unit for ovalbumin is 7.7 kilobases. The DNA sequence of the region surrounding the 5' end of the ovalbumin gene is presented. Comparison of this sequence with those of other eukaryotic genes reveals striking similarities, possibly related to a promoter region, approximately 30 base pairs upstream from the site coding for the 5' end of the mRNA.