Showing papers on "Gene published in 1984"
TL;DR: The complete (172,282 base pairs) nucleotide sequence of the B95-8 strain of Epstein–Barr virus has been established using the dideoxynucleotide/M13 sequencing procedure.
Abstract: The complete (172,282 base pairs) nucleotide sequence of the B95-8 strain of Epstein-Barr virus has been established using the dideoxynucleotide/M13 sequencing procedure. Many RNA polymerase II promoters have been mapped and the mRNAs from these promoters have been assigned to the latent or early/late productive virus cycles. Likely protein-coding regions have been identified and three of these have been shown to encode a ribonucleotide reductase, a DNA polymerase and two surface glycoproteins.
2,016 citations
TL;DR: A new method for in vitro insertional mutagenesis of genes cloned in Escherichia coli that makes use of the omega fragment, a 2.0-kb DNA segment consisting of an antibiotic resistance gene flanked by short inverted repeats carrying transcription and translation termination signals and synthetic polylinkers.
1,729 citations
TL;DR: Many, probably even all possible types of simple sequence are repetitive components of eukaryotic genomes and it is proposed that they arise by common mechanisms namely slippage replication and unequal crossover and that they might have no general function with regards to gene expression.
Abstract: Simple sequences are stretches of DNA which consist of only one, or a few tandemly repeated nucleotides, for example poly (dA) X poly (dT) or poly (dG-dT) X poly (dC-dA). These two types of simple sequence have been shown to be repetitive and interspersed in many eukaryotic genomes. Several other types have been found by sequencing eukaryotic DNA. In this report we have undertaken a systematical survey for simple sequences. We hybridized synthetical simple sequence DNA to genome blots of phylogenetically different organisms. We found that many, probably even all possible types of simple sequence are repetitive components of eukaryotic genomes. We propose therefore that they arise by common mechanisms namely slippage replication and unequal crossover and that they might have no general function with regards to gene expression. This latter inference is supported by the fact that we have detected simple sequences only in the metabolically inactive micronucleus of the protozoan Stylonychia, but not in the metabolically active macronucleus which is derived from the micronucleus by chromosome diminution.
1,382 citations
TL;DR: Stimulation of fibroblasts with serum or purified growth factors leads to a dramatic induction of expression of both c-fos mRNA and protein within a few minutes, followed by activation of c-myc.
Abstract: Stimulation of fibroblasts with serum or purified growth factors leads to a dramatic induction of expression of both c-fos mRNA and protein within a few minutes, followed by activation of c-myc. This suggests that c-fos induction is a primary event and the earliest known effect on gene expression by growth factors.
1,283 citations
TL;DR: A series of rat neuro/glioblastomas all contain the same transforming gene (neu) which induces synthesis of a tumour antigen of relative molecular mass (Mr) 185,000 (p185), which is serologically related to the epidermal growth factor (EGF) receptor.
Abstract: A series of rat neuro/glioblastomas all contain the same transforming gene (neu) which induces synthesis of a tumour antigen of relative molecular mass (Mr) 185,000 (p185). The neu oncogene bears homology to erb-B and the tumour antigen, p185, is serologically related to the epidermal growth factor (EGF) receptor. The two proteins, EGF receptor and p185 appear to be distinct, as they coexist in nontransformed Rat-1 cells.
1,158 citations
TL;DR: The complete 186,000 base-pair (bp) human factor VIII gene has been isolated and consists of 26 exons ranging in size from 69 to 3,106 bp and introns as large as 32.4 kilobases as mentioned in this paper.
Abstract: The complete 186,000 base-pair (bp) human factor VIII gene has been isolated and consists of 26 exons ranging in size from 69 to 3,106 bp and introns as large as 32.4 kilobases (kb). Nine kb of mRNA and protein-coding DNA has been sequenced and the mRNA termini have been mapped. The relationship between internal duplications in factor VIII and evolution of the gene is discussed.
988 citations
TL;DR: A repetitive DNA sequence has been identified in the Drosophila melanogaster genome that appears to be localized specifically within genes of the bithorax and Antennapedia complexes that are required for correct segmental development.
Abstract: A repetitive DNA sequence has been identified in the Drosophila melanogaster genome that appears to be localized specifically within genes of the bithorax and Antennapedia complexes that are required for correct segmental development. Initially identified in cloned copies of the genes Antennapedia, Ultrabithorax and fushi tarazu, the sequence is also contained within two other DNA clones that have characteristics strongly suggesting that they derive from other homoeotic genes.
968 citations
TL;DR: Three mutants of Arabidopsis thaliana (L.) Heynh were characterized by a reduced seed dormancy and by symptoms of withering, indicating disturbed water relations and resembled phenotypically the ABA-deficient mutants the authors described earlier in this species.
Abstract: Abscisic acid (ABA) insensitive mutants of Arabidopsis thaliana (L.) Heynh. were isolated by selecting plants which grew well on a medium containing 10 μM ABA. From the progeny of approximately 3500 mutagen-treated seeds, five mutants of at least three different loci were isolated. Three mutants were characterized, moreover, by a reduced seed dormancy and by symptoms of withering, indicating disturbed water relations and, therefore, resembled phenotypically the ABA-deficient mutants we described earlier in this species. Two mutants showed in addition only a reduction of seed dormancy. Compared to wild type, all mutants showed similar or increased levels of endogenous ABA in developing seeds and fruits (siliquae). The role of the different genes involved is discussed in relation to the mechanism of ABA action.
954 citations
TL;DR: Molecular cloning of the transforming gene from a chemically transformed human osteosarcoma-derived cell line enables the gene to be mapped to chromosome 7 (7p11.4–7qter) and by direct hybridization to be shown to be unrelated to known oncogenes.
Abstract: Molecular cloning of the transforming gene from a chemically transformed human osteosarcoma-derived cell line enables the gene to be mapped to chromosome 7 (7p11.4-7qter) and by this criterion and by direct hybridization to be shown to be unrelated to known oncogenes.
951 citations
TL;DR: A mathematical formula for estimating the average number of nucleotide substitutions per site (6) between two homologous DNA sequences is developed by taking into account unequal rates of substitution among different nucleotide pairs as discussed by the authors.
Abstract: A mathematical formula for estimating the average number of nucleotide substitutions per site (6) between two homologous DNA sequences is developed by taking into account unequal rates of substitution among different nucleotide pairs. Although this formula is obtained for the equal-input model of nucleotide substitution, computer simulations have shown that it gives a reasonably good estimate for a wide range of nucleotide substitution patterns as long as 6 is equal to or smaller than 1. Furthermore, the frequency of cases to which the formula is inapplicable is much lower than that for other similar methods recently proposed. This point is illustrated using insulin genes. A statistical method for estimating the number of nucleotide changes due to deletion and insertion is also developed. Application of this method to globin gene data indicates that the number of nucleotide changes per site increases with evolutionary time but the pattern of the increase is quite irregular.
TL;DR: It is found that there is a B-cell-specific enhancer-activator region in the J-C intron of the human kappa gene that is necessary for efficient transcription of the cloned gene in mouse pre-B lymphocytes, suggesting that both the DNA element and the proteins required for its regulatory activity have been highly conserved in evolution.
Abstract: We have developed a general method for introducing cloned genes into mammalian cells that affords substantial benefits over current technology. It is simple, rapid, and applicable to many (perhaps all) cell types, including those that are refractory to traditional transfection procedures. The method involves exposure of a suspension of cells and cloned DNA to a high-voltage electric discharge. In a model application of this transfection procedure, we have studied the expression of cloned human and mouse Ig kappa genes stably introduced into mouse pre-B cells and fibroblasts. We find that there is a B-cell-specific enhancer-activator region in the J-C intron of the human kappa gene that is necessary for efficient transcription of the cloned gene in mouse pre-B lymphocytes. This suggests that both the DNA element and the proteins required for its regulatory activity have been highly conserved in evolution and that these elements operate at the pre-B-cell stage of immunocyte development, a stage that precedes productive kappa gene rearrangement.
TL;DR: The GAL1 and GAL10 genes of Saccharomyces cerevisiae are divergently transcribed, with 606 base pairs of DNA separating their transcription initiation sites as mentioned in this paper.
Abstract: The GAL1 and GAL10 genes of Saccharomyces cerevisiae are divergently transcribed, with 606 base pairs of DNA separating their transcription initiation sites. These two genes are stringently coregulated: their expression is induced ca. 1,000-fold in cells growing on galactose and is repressed by growth on glucose. The nucleotide sequence of the region of DNA between these genes and the precise sites of transcription initiation are presented here. The most notable feature of the nucleotide sequence of this region is a 108-base-pair guanine-plus-cytosine-rich stretch of DNA located approximately in the middle of the region between GAL1 and GAL10. Analysis of the effects of mutations that alter the region between these two genes, constructed in vitro or selected in vivo, suggest that these guanine-plus-cytosine-rich sequences are required for the expression of both genes. The region of DNA between GAL1 and GAL10 is sufficient for regulation of expression of these genes: fusion of the region to the yeast HIS3 gene places HIS3 under GAL control.
TL;DR: Two stretches of DNA (genetic elements), lying close to the promoter for a human gene for metallothionein, that separately mediate the induction of the gene by heavy metal ions, particularly cadmium, and by glucocorticoid hormones are defined.
Abstract: Deletion experiments have defined two stretches of DNA (genetic elements), lying close to the promoter for a human gene for metallothionein, that separately mediate the induction of the gene by heavy metal ions, particularly cadmium, and by glucocorticoid hormones. The element responsible for induction by cadmium is duplicated, yet a single copy is fully functional; the element responsible for induction by glucocorticoid hormones is coincident with the DNA-binding site for the glucocorticoid hormone receptor.
Patent•
15 Aug 1984
TL;DR: A method for detecting the presence of the gene for Huntington's disease in a subject which comprises analyzing the human chromosome 4 of the subject for a DNA polymorphism, preferably a restriction fragment length polymorphism (RFLP), linked to Huntington's Disease was proposed in this paper.
Abstract: A method for detecting the presence of the gene for Huntington's disease in a subject which comprises analyzing the human chromosome 4 of the subject for a DNA polymorphism, preferably a restriction fragment length polymorphism (RFLP), linked to Huntington's Disease.
TL;DR: This finding has been exploited to identify the coding sequence of the viomycin phosphotransferase gene of Streptomyces vinaceus, and an easily applied computer program has been written to carry out and display such analyses.
TL;DR: Overall, this transfection protocol using DEAE-dextran coupled to a shock treatment is simple, straightforward, and gives consistently high levels of expression of the input DNA.
Abstract: Using a plasmid containing the bacterial chloramphenicol acetyl transferase gene, we have assayed for transient expression of DNA introduced into mouse L cells by a variety of transfection conditions. High efficiency uptake and expression of this foreign DNA have been achieved by modifying the DEAE dextran mediated transfection procedure of McCutchan and Pagano (1) to include a shock with either dimethyl sulfoxide or glycerol. Inclusion of the shock step can increase expression of the transfected gene a surprising approximately 50 fold. With plasmid constructs that do not replicate after transfection, we can readily detect CAT activity in an overnight autoradiographic exposure from less than 0.1% of an extract from a 60 mm dish of transfected cells. We have determined the amounts of DNA, the amount and time course of DEAE-dextran and dimethyl sulfoxide treatments, the effects of additional DNA, and the time after transfection which yield maximal expression. Overall, this transfection protocol using DEAE-dextran coupled to a shock treatment is simple, straightforward, and gives consistently high levels of expression of the input DNA.
TL;DR: The genotype at this locus of 393 unrelated diabetic and nondiabetic individuals is determined and differences were observed in the genotypie and allelic frequencies between groups of different races.
Abstract: A polymorphic region flanking the human insulin gene on the short arm of chromosome 11, the insulin-gene-linked DNA polymorphism, can be described as a locus with at least three classes of alleles: a common small “class 1” allele averaging 570 base pairs, a rare intermediate “class 2” allele of about 1320 base pairs, and a large “class 3” allele averaging 2470 base pairs in size.
We have determined the genotype at this locus of 393 unrelated diabetic and nondiabetic individuals. Differences were observed in the genotypie and allelic frequencies between groups of different races. Asians [17 nondiabetic, 2 with insulin-dependent diabetes mellitus (IDDM), and 8 with non-insulin-dependent diabetes mellitus (NIDDM)] exhibited the least variation in the size of this locus and 98% of the alleles in this group were class 1. A group of American blacks (32 nondiabetic, 5 with IDDM, and 40 with NIDDM) exhibited considerable variation in the size of this locus, and about 22% of the individuals examined had a genotype that included a rare class 2 allele. In neither of these two racial groups were the genotypie or allelic frequencies different between the nondiabetic and diabetic segments of these groups.
However, in a group of Caucasians (83 nondiabetic, 113 with IDDM, and 76 with NIDDM), there was a significantly higher frequency of class 1 alleles and genotypes containing two class 1 alleles in the diabetic patients compared with nondiabetic controls. A strikingly higher frequency of class 1 alleles (P < 0.0001) and genotypes containing two class 1 alleles (P < 0.0001) was observed in the IDDM group compared with nondiabetic Caucasians, whereas the difference between NIDDM and controls was only near the level of statistical significance (P = 0.025). Analysis of the combined data of our Caucasian population and those reported from studies of Caucasians from Denmark and St. Louis, Missouri, continued to show an increased frequency of class 1 alleles and genotypes containing two class 1 alleles in the IDDM group (P = 0.0001) compared with the nondiabetic group; however, there were no longer differences in genotypic or allelic frequencies between NIDDM and nondiabetic groups.
TL;DR: The results indicate that the p53-encoding gene can play a causal role in the conversion of normal fibroblasts into tumorigenic cells, suggesting an important role of p53 in tumorigenesis.
Abstract: The protein p53 is highly expressed in a large variety of transformed cell types originating from diverse species. These include cells transformed by Simian virus 40 (SV40), adenovirus and Abelson virus, as well as a variety of chemically transformed cells. Substantial amounts of p53 are also present in certain non-transformed cells, for example, some embryonic tissues. The protein may be localized in different cellular compartments in normal and transformed cells. The strong correlation between tumorigenicity and high levels of p53 suggests an important role of p53 in tumorigenesis. We report here experiments in which we have co-transfected the murine cellular gene encoding for p53 with a ras gene into primary rat embryo fibroblasts. Our results indicate that the p53-encoding gene can play a causal role in the conversion of normal fibroblasts into tumorigenic cells.
TL;DR: The gene for the circumsporozoite (CS) protein of Plasmodium falciparum has been cloned and its nucleotide sequence determined and suggests that they may have a function such as binding to liver cells and may represent an invariant target for immunity.
Abstract: The gene for the circumsporozoite (CS) protein of Plasmodium falciparum has been cloned and its nucleotide sequence determined. The gene encodes a protein of 412 amino acids as deduced from the nucleotide sequence. The protein contains 41 tandem repeats of a tetrapeptide, 37 of which are Asn-Ala-Asn-Pro and four of which are Asn-Val-Asp-Pro. Monoclonal antibodies against the CS protein of Plasmodium falciparum were inhibited from binding to the protein by synthetic peptides of the repeat sequence. The CS protein of Plasmodium falciparum and the CS protein of a simian malaria parasite, Plasmodium knowlesi, have two regions of homology, one of which is present on either side of the repeat. One region contains 12 of 13 identical amino acids. Within the nucleotide sequence of this region, 25 of 27 nucleotides are conserved. The conservation of these regions in parasites widely separated in evolution suggests that they may have a function such as binding to liver cells and may represent an invariant target for immunity.
TL;DR: Plants regenerated from transformed cell lines were phenotypically normal and fertile, and they maintained and expressed the foreign gene throughout the development of vegetative and generative organs.
Abstract: Evidence for direct, gene-mediated stable genetic transformation of plant cells of Nicotiana tabacum is presented. A selectable hybrid gene comprising the protein coding region of the Tn5 aminoglycoside phosphotransferase type II gene under control of cauliflower mosaic virus gene VI expression signals was introduced into plant protoplasts as part of an Escherichia coli plasmid. The gene was stably integrated into plant genomic DNA and constitutively expressed in selected, drug resistant, protoplast-derived cell clones. The mode of integration of the foreign gene into the plant genome resembled that observed for DNA transfection of mammalian cells. Plants regenerated from transformed cell lines were phenotypically normal and fertile, and they maintained and expressed the foreign gene throughout the development of vegetative and generative organs. Microspores, grown in anther culture, developed into resistant and sensitive haploid plantlets. Genetic crossing analysis of one of the transformed plants revealed the presence of one dominant trait for kanamycin resistance segregating in a Mendelian fashion in the F(1) generation.
TL;DR: Gene dosage effects suggest that KEX2 is the structural gene for the endopeptidase, and cloned DNA restores both enzymatic activity in vitro and the normal pattern of proteolytic processing and glycosylation of prepro-alpha-factor in vivo.
TL;DR: Sequence analysis of the cross-hybridizing DNA from the three loci revealed the conservation of predicted amino acid sequences derived from coding parts of the genes, suggesting that two homoeotic loci and a "segment-deficient" locus encode protein products with partially shared structures and that theThree loci may be evolutionarily and functionally related.
Abstract: Genes that regulate the development of the fruit fly Drosophila melanogaster exist as tightly linked clusters in at least two cases. These clusters, the bithorax complex (BX-C) and the Antennapedia complex (ANT-C), both contain multiple homoeotic loci: mutations in each locus cause a transformation of one part of the fly into another. Several repetitive DNA sequences, including at least one transposon, were mapped in the ANT-C. DNA from the 3' exon of Antennapedia (Antp), a homoeotic locus in the ANT-C, hybridized weakly to DNA from the 3' exon of Ultrabithorax (Ubx), a homoeotic locus in the BX-C. DNA from each of these 3' exons also hybridized weakly to DNA from the fushi tarazu locus of the ANT-C. The fushi tarazu (ftz) locus controls the number and differentiation of segments in the developing embryo. Sequence analysis of the cross-hybridizing DNA from the three loci revealed the conservation of predicted amino acid sequences derived from coding parts of the genes. This suggests that two homoeotic loci and a "segment-deficient" locus encode protein products with partially shared structures and that the three loci may be evolutionarily and functionally related.
TL;DR: Comparison of the sequence of a cloned T cell-specific cDNA with those of cross-reacting cloned cDNAs isolated from a thymocyte library indicates the presence of variable, constant and joining regions remarkably similar in size and sequence to those encoding immunoglobulin proteins.
Abstract: Comparison of the sequence of a cloned T cell-specific cDNA with those of cross-reacting cloned cDNAs isolated from a thymocyte library indicates the presence of variable, constant and joining regions remarkably similar in size and sequence to those encoding immunoglobulin proteins. Together with the evidence for somatic gene rearrangements reported in the accompanying paper, this strongly suggests that the TM86 cDNA clone encodes one chain of the T-cell receptor for antigen.
TL;DR: The chromosomal breakpoint of chronic lymphocytic leukemia (CLL) cells of the B-cell type carrying the translocated long arms of chromosomes 11 and 14 [t(11;14) (q13;q32)] was cloned and a gene, named bcl -1, appears to be unrelated to any of the known retrovirus oncogenes described to date.
Abstract: The chromosomal breakpoint of chronic lymphocytic leukemia (CLL) cells of the B-cell type carrying the translocated long arms of chromosomes 11 and 14 [t(11;14) (q13;q32)] was cloned. The breakpoint was found to be within the joining segment of the human heavy chain locus on the translocated long arm of chromosome 14. A probe that is specific for chromosome 11 and that maps immediately 5' to the breakpoint on the 14q+ chromosome was isolated. The probe detected a rearrangement of the homologous genomic DNA segment in the parental CLL cells and also in DNA from a diffuse large cell lymphoma with the t(11;14) translocation. This rearranged DNA segment was not present in Burkitt lymphoma cells with the t(8;14) translocation or in nonneoplastic human lymphoblastoid cells. The probe can thus be used to identify and characterize a gene located on band q13 of chromosome 11 that appears to be involved in the malignant transformation of human B cells carrying the t(11;14) translocation. This gene, named bcl -1, appears to be unrelated to any of the known retrovirus oncogenes described to date.
TL;DR: A genetic analysis was reported to identify the luminescence genes (lux) that reside on this recombinant plasmid and the corresponding gene products were identified from the complementation analysis and the minicell programing experiments.
Abstract: Expression of luminescence in Escherichia coli was recently achieved by cloning genes from the marine bacterium Vibrio fischeri. One DNA fragment on a hybrid plasmid encoded regulatory functions and enzymatic activities necessary for light production. We report the results of a genetic analysis to identify the luminescence genes (lux) that reside on this recombinant plasmid. lux gene mutations were generated by hydroxylamine treatment, and these mutations were ordered on a linear map by complementation in trans with a series of polar transposon insertions on other plasmids. lux genes were defined by complementation of lux gene defects on pairs of plasmids in trans in E. coli. Hybrid plasmids were also used to direct the synthesis of polypeptides in the E. coli minicell system. Seven lux genes and the corresponding gene products were identified from the complementation analysis and the minicell programing experiments. These genes, in the order of their position on a linear map, and the apparent molecular weights of the gene products are luxR (27,000), luxI (25,000), luxC (53,000), luxD (33,000), luxA (40,000), luxB (38,000), and luxE (42,000). From the luminescence phenotypes of E. coli containing mutant plasmids, functions were assigned to these genes: luxA, luxB, luxC, luxD, and luxE encode enzymes for light production and luxR and luxI encode regulatory functions.
TL;DR: It is proposed that the micRNA inhibits the translation of the ompF mRNA by hybridizing with it and this RNA interaction may cause premature termination of the transcription of theOmpF gene or destabilization of the OmpF RNA or both.
Abstract: The expression of the genes for the major outer membrane proteins OmpF and OmpC are osmoregulated. The ompC locus was found to be transcribed bidirectionally under conditions of high osmolarity and a 174-base transcript encoded upstream of ompC was found to inhibit the OmpF production and to substantially reduce the amount of the ompF mRNA. This RNA [mRNA-interfering complementary RNA (micRNA)] has a long sequence that is complementary to the 5' end region of the ompF mRNA. We propose that the micRNA inhibits the translation of the ompF mRNA by hybridizing with it. This RNA interaction may cause premature termination of the transcription of the ompF gene or destabilization of the ompF mRNA or both.
TL;DR: A number of mink cell focus-forming (MCF) proviruses was molecularly cloned from mouse lymphoma DNA to detect common integration regions in other MuLV-induced lymphomas and variations in the molecular structure of the corresponding region (Pim-1) in other lymphomas were revealed.
TL;DR: The molecular biology of a new class of genes, called osm (osmotic tolerance) genes, that protect bacteria like Escherichia coli against osmotic stress and may work in a similar manner in plants and animals are concerned.
Abstract: The drought of 1983 resulted in some 10 billion dollars in agricultural losses and has focused attention on the vulnerability of our major crops to this devastating form of environmental stress. This article is concerned with the molecular biology of a new class of genes, called osm (osmotic tolerance) genes, that protect bacteria like Escherichia coli against osmotic stress and may work in a similar manner in plants and animals. Osm genes govern the production of a class of molecules, such as betaine and proline, that protect the cell and its constituents against dehydration. These osmoprotectant molecules have been known for many years to accumulate in plants but have only recently been shown to have potent antistress activity for bacteria.