Showing papers on "Gene published in 1980"
TL;DR: The results demonstrate that genes can be introduced into the mouse genome by direct insertion into the nuclei of early embryos by microinjected into pronuclei of fertilized mouse oocytes.
Abstract: A recombinant plasmid composed of segments of herpes simplex virus and simian virus 40 viral DNA inserted into the bacterial plasmid pBR322 was microinjected into pronuclei of fertilized mouse oocytes. The embryos were implanted in the oviducts of pseudopregnant females and allowed to develop to term. DNA from newborn mice was evaluated by the Southern blotting technique for the presence of DNA homologous to the injected plasmid. Two of 78 mice in one series of injections showed clear homology, though the injected sequences had been rearranged. Band intensities from the two positive mice were consistent with the presence of donor DNA in most or all of the cells of the newborns. These results demonstrate that genes can be introduced into the mouse genome by direct insertion into the nuclei of early embryos. This technique affords the opportunity to study problems of gene regulation and cell differentiation in a mammalian system by application of recombinant DNA technology.
1,417 citations
TL;DR: A large number of the ribosomal genes affected by the bobbed locus mutation in Drosophila belong to the TSP class, which has been associated with central giant cell reprograming.
Abstract: PERSPECTIVES AND SUMMARY ,. '728 RIBOSOMAL RNA GENES 729 Introduction 729 Ribosomal Gene Redundancy 730 Chromosomal Location of Ribosomal Genes 730 Structure of Ribosomal Genes 735 Integrated tandemly repeated ribosomal genes 738 Extrachromosomal rDNA 741 Changes in Redundancy of Ribosomal Genes 741 Amplification of ribosomal genes 742 Mutants of the bobbed locus in Drosophila 743 THE GENES FOR 5S RIBOSOMAL RNA 745 tRNA GENES 748 PROPERTIES OF SPACERS 749 GENE FAMILIES WITH VARIANT REPETITION 751 Globin genes 751 Actin genes 751 Immunoglobulin genes 752 Vitel/ogenin genes 753 Ovalbumin and genes X and Y 753 Chorion protein genes 753
1,286 citations
TL;DR: A model for the involvement of short direct repeat sequences in the generation of deletions in the noncoding and coding regions of B-like globin genes during evolution is described.
1,097 citations
TL;DR: Direct microinjection of DNA by glass micropipettes was used to introduce the Herpes simplex virus thymidine kinase gene into cultured mammalian cells, and transformation frequency was relatively insensitive to DNA concentration and did not depend on co-injecting with a carrier DNA.
1,045 citations
TL;DR: A series of plasmid vectors suitable for the detection and cloning of translational control signals and 5' coding sequences of exogenously derived genes yielded hybrid proteins useful for purifying peptide sequences encoded by exogenous deoxyribonucleic acid fragments and for studies relating the structure and function of specific peptide segments.
Abstract: We report the construction and use of a series of plasmid vectors suitable for the detection and cloning of translational control signals and 5' coding sequences of exogenously derived genes. In these plasmids, the first eight codons of the amino-terminal end of the lactose operon beta-galactosidase gene, lacZ, were removed, and unique BamHI, EcoRI, and SmaI (XmaI) endonuclease cleavage sites were incorporated adjacent to the eighth codon of lacZ. Introduction of deoxyribonucleic acid fragments containing appropriate regulatory signals and 5' coding sequences into such lac fusion plasmids led to the production of hybrid proteins consisting of the carboxyl-terminal segment of a beta-galactosidase remnant plus a peptide fragment that contained the amino-terminal amino acids encoded by the exogenous deoxyribonucleic acid sequence. These hybrid peptides retained beta-galactosidase enzymatic activity and yielded a Lac+ phenotype. Such hybrid proteins are useful for purifying peptide sequences encoded by exogenous deoxyribonucleic acid fragments and for studies relating the structure and function of specific peptide segments.
971 citations
TL;DR: The sequence data suggest that intergenic conversions occur in the germ line, and strongly suggest that DNA sequence polymorphisms for localized deletions, additions and base substitutions are very common in human populations.
810 citations
TL;DR: The sequence of regions of the chicken ovalbumin gene believed to be important in the control of initiation of transcription, splicing, and transcription termination or polyadenylation are presented.
Abstract: We present the sequence of regions of the chicken ovalbumin gene believed to be important in the control of initiation of transcription, splicing, and transcription termination or polyadenylation. Comparison with corresponding areas of other genes reveals some homologous regions which might play a role in these processes.
758 citations
TL;DR: A model for variable region gene rearrangement mediated by proteins which recognize the same conserved sequences adjacent to both light and heavy chain immunoglobulin gene segments is proposed.
736 citations
TL;DR: Transfection of cultured monkey kidney cells with recombinant DNA constructed with a cloned Escherichia coli gene that codes for xanthine-guanine phosphoribosyltransferase and several different SV40 DNA-based vectors results in the synthesis of readily measurable quantities of the bacterial enzyme.
Abstract: Transfection of cultured monkey kidney cells with recombinant DNA constructed with a cloned Escherichia coli gene that codes for xanthine-guanine phosphoribosyltransferase and several different SV40 DNA-based vectors, results in the synthesis of readily measurable quantities of the bacterial enzyme. Moreover, the physiological defect in purine nucleotide synthesis characteristic of human Lesch-Nyhan cells can be overcome by the introduction of the bacterial gene into these cells.
724 citations
TL;DR: Two types of somatic recombination are necessary for the generation of a complete immunoglobulin γ2b gene from germ-line DNA sequences and DNA sequencing studies suggest that the two types of recombination operate by different mechanisms.
Abstract: At least two types of somatic recombination are necessary for the generation of a complete immunoglobulin gamma 2b gene from germ-line DNA sequences. The first type of recombination consists of the assembly of three separate DNA segments, each encoding a different part of the variable region. The second type of recombination replaces the exons coding for the constant region of the mu chain with those coding for the same region of the gamma 2b chain. The DNA sequencing studies suggest that the two types of recombination operate by different mechanisms.
690 citations
TL;DR: Examination of the DNA from seven members of a family revealed fragment lengths that are consistent with their inheritance as Mendelian alleles through three generations, and appears to be the result of DNA rearrangements rather than base-pair substitutions or modifications.
Abstract: A locus in the human genome, not associated with any specific gene, has been found to be a site of restriction fragment length polymorphism. The polymorphism was found by hybridizing a 16-kilobase-pair segment of single-copy human DNA, selected from the human genome library cloned in phage lambda CH4A, to a Southern transfer of total human DNA digested with EcoRI. DNAs from a number of individuals from within Mormon pedigrees as well as random individuals have been examined. The locus is highly variable, with at least eight alleles present, homozygotes accounting for less than 25% of the individuals examined. The polymorphism appears to be the result of DNA rearrangements rather than base-pair substitutions or modifications. Examination of the DNA from seven members of a family revealed fragment lengths that are consistent with their inheritance as Mendelian alleles through three generations.
TL;DR: It is concluded that not all of these sites are neutral and that they do not behave as accurate evolutionary clocks over long periods of time, but nucleotide substitutions leading to amino acid replacements are an excellent clock.
TL;DR: The DNA sequence of a 1.45 kb EcoRI fragment from the yeast (Saccharomyces cerevisiae) TRP1 region has been determined and theTRP1 gene has been located on the fragment by analysis of potential initiation and termination codons in the DNA sequence.
TL;DR: Analysis of DNA modification in the human γδβ-globin gene region suggests that a low level of DNA methylation may be a necessary, but not a sufficient, condition for gene expression in higher eucaryotes.
TL;DR: A comparison of the human with the rat insulin genes indicates potential regulatory regions in the DNA segment preceding the gene and suggests that the ancestral form of the insulin gene had two intervening sequences.
Abstract: The human insulin gene contains two intervening sequences, one is within the region transcribed into the 5'-untranslated segment of the mRNA and the other interrupts the C-peptide encoding region. A comparison of the human with the rat insulin genes indicates potential regulatory regions in the DNA segment preceding the gene and suggests that the ancestral form of the insulin gene had two intervening sequences.
TL;DR: Concerted evolution appears also to have occurred, but far more slowly, in the region coding for the adult beta-like chains of hemoglobin, leading to the hypothesis that the lengths of the noncoding regions are important determinants of the rates at which genes are gained and lost by intergenic recombination.
Abstract: Rapid cycles of gene duplication and loss appear to have been going on in the region coding for the alpha chain of adult hemoglobin. This is inferred from restriction endonuclease analysis of the alpha gene region in five species of apes, whose common ancestor lived about 10 million years ago. Because all five species resemble humans in having duplicate alpha genes, the duplicate state of this region is probably at least as old as the common ancestor of all these species. However, the alpha polypeptides within these species are about 10 times more alike than is expected for 10 million years of divergent evolution. Thus, the alpha polypeptides within each species have been evolving in concert. Changes in gene number have also taken place in the apes. Whereas the predominant number of alpha genes per chromosome is two for most species, it is three for chimpanzees. Concerted evolution appears also to have occurred, but far more slowly, in the region coding for the adult beta-like chains of hemoglobin. Consideration of the structural differences between the two regions leads to the hypothesis that the lengths of the noncoding regions are important determinants of the rates at which genes are gained and lost by intergenic recombination.
TL;DR: Results indicate that E. coli contains a set of genes that, like prophage lambda genes, are expressed in response to DNA-damaging agents and regulated by the recA and lexA gene products.
Abstract: Operon fusions in Escherichia coli were obtained that showed increased beta-galactosidase expression in response to treatment with the DNA-damaging agent mitomycin C. These fusions were generated by using the Mud(ApR, lac) vector [Casadaban, M.J. & Cohen, S.N. (1979) Proc. Natl. Acad. Sci. USA 76, 4530-4533] to insert the lactose structural genes randomly into the bacterial chromosome. Induction of beta-galactosidase in these strains, which carried fusions of lac to these din (damage-inducible) loci, was (i) triggered by UV light as well as by mitomycin C and (ii) abolished by either a recA- or a lexA- mutation. Similar characteristics of induction were observed when the lactose genes were fused to a prophage lambda promoter by using Mud(ApR, lac). These results indicate that E. coli contains a set of genes that, like prophage lambda genes, are expressed in response to DNA-damaging agents and regulated by the recA and lexA gene products. These din genes map at five bacterial loci. One din::Mud(ApR, lac) insertion results in a UV-sensitive phenotype and may be within the uvrA transcriptional unit.
TL;DR: The results of these experiments indicate that the tK gene is not interrupted by intervening DNA sequences, and that certain oligonucleotide sequences adjacent to the termini of the tk gene are homologous to similarly positioned sequences common to structural genes of eukaryotic cells.
Abstract: This paper presents the nucleotide sequence of the Herpes Simplex Virus thymidine kinase (tk) gene. The position on the DNA sequence corresponding to the 5' and 3' termini of tk messenger RNA have been mapped. The mRNA termini are separated by slightly more than 1,300 nucleotides. The same 2,300 nucleotide segment of tk coding strand DNA is fully protected from S1 nuclease digestion when hybridized to tk mRNA. The location and size of the mRNA-coding segment corresponds to a region of the viral DNA that is essential for tk gene expression in microinjected frog oocytes. The nucleotide sequence of the HSV tk gene exhibits an open translational reading frame of 376 codons that extends from the methionine codon most proximal to the 5' terminus of tk mRNA to a UGA stop codon approximately 70 nucleotides from the poly-A addition site. The results of these experiments indicate that the tk gene is not interrupted by intervening DNA sequences, and that certain oligonucleotide sequences adjacent to the termini of the tk gene are homologous to similarly positioned sequences common to structural genes of eukaryotic cells.
TL;DR: The alcohol dehydrogenase (ADH) gene (Adh) of Drosophila melanogaster was isolated by utilizing a mutant strain in which the Adh locus is deleted, and two intervening sequences were identified within Adh by S1 nuclease mapping experiments.
Abstract: The alcohol dehydrogenase (ADH; alcohol: NAD+ oxidoreductase, EC 1.1.1.1) gene (Adh) of Drosophila melanogaster was isolated by utilizing a mutant strain in which the Adh locus is deleted. Adult RNA from wild-type flies was enriched in ADH sequences by gel electrophoresis and then used to prepare labeled cDNA for screening a bacteriophage lambda library of genomic Drosophila DNA. Of the clones that hybridized in the initial screen, one clone was identified that hybridized with labeled cDNA prepared from a wild-type Drosophila strain but did not hybridize with cDNA prepared from an Adh deletion strain. This clone was shown to contain ADH structural gene sequences by three criteria: in situ hybridization, in vitro translation of mRNA selected by hybridization to the cloned DNA, and comparison of the ADH protein sequence with a nucleotide sequence derived from the cloned DNA. Comparison of the restriction site maps from clones of three different wild-type Drosophila strains revealed the presence of a 200-nucleotide sequence in one strain that was absent from the other two strains. The ADH mRNA sequences were located within the cloned DNA by hybridization mapping experiments. Two intervening sequences were identified within Adh by S1 nuclease mapping experiments.
TL;DR: In this paper, the authors used cloned actin sequences from Dictyostelium discoideum to identify and clone the actin gene in yeast and determined the nucleotide sequence of that gene and its flanking regions.
Abstract: The yeast Saccharomyces cerevisiae is known to contain the highly conserved and unbiquitous protein actin. We have used cloned actin sequences from Dictyostelium discoideum to identify and clone the actin gene in yeast. Hybridization to genomic fragments of yeast DNA suggest that there is a single actin gene in yeast. We have determined the nucleotide sequence of that gene and its flanking regions. The sequence of the gene reveals an intervening sequence of 309 base pairs in the coding sequences at the 5' end of the gene. The existence and location of the intervening sequence was verified by using the dideoxy chain termination technique to determine the sequence at the 5' terminus of the actin mRNA. The similarity of the splice junction sequences in this gene to those found in higher eukaryotes suggests that yeast must possess a similar splicing enzyme.
TL;DR: The nucleotide sequence of the lacY gene coding for lactose permease (M protein) in Escherichia coli has been determined and is predicted to consist of 417 residues (71% nonpolar), resulting in a protein with a molecular weight of 46,504.
Abstract: The nucleotide sequence of the lacY gene coding for lactose permease (M protein) in Escherichia coli has been determined. The sequence includes the intergenic regions between the lacZ (β-galactosidase) and lacY genes as well as the region between the lacY and lacA (transacetylase) genes. Lactose permease is predicted to consist of 417 residues (71% nonpolar), resulting in a protein with a molecular weight of 46,504. The reading frame was confirmed by the sequence of a nonsense mutation changing codon 33 from UGG to UAG.
TL;DR: It is found that human and ape ribosomal genes undergo concerted evolution involving genetic exchanges among nucleolus organizers on nonhomologous chromosomes.
Abstract: We have found that human and ape ribosomal genes undergo concerted evolution involving genetic exchanges among nucleolus organizers on nonhomologous chromosomes. This conclusion is based upon restriction enzyme analysis of the ribosomal gene families in man and five ape species. Certain structural features were found to differ among (but not within) species even though the ribosomal genes have a multichromosomal distribution. Genetic exchanges among nucleolus organizer regions may be related to the well-known phenomenon of acrocentric chromosome associations observed in man and apes. Length variation in a region of the nontranscribed spacer was found in both chimpanzee species we examined. The nature of this length variation was found to be identical to that previously described in man. The origin of the length variation and its polymorphism within these three species might be explained by unequal alignment and unequal cross-over among the ribosomal genes. An especially surprising finding was a nucleotide sequence polymorphism present in each individual human and ape we examined. Some ribosomal genes of each individual have a HindII site in the 28S gene about 800 base pairs from the EcoRI site in this gene. The remaining 28S genes lack this HindII site. The presence of this polymorphism within individuals of every species we examined suggests that it has been maintained by natural selection.
TL;DR: Analysis of the cloned DNA confirms the linkage arrangement of the two adult α-globin genes (α1 and α2) previously derived from genomic blotting experiments and identifies two additional closely linked α-like genes.
TL;DR: The complete nucleotide sequence of the human β-globin gene is reported to provide the basis for comparing normal β- globin genes with those obtained from the DNA of individuals with genetic defects in hemoglobin expression.
TL;DR: The 12 Interferon (IFN)-related sequences detected in a human gene bank fall into not less than eight distinct classes, indicating that there are at least eight IFN-related genes.
Abstract: The 12 Interferon (IFN)-related sequences detected in a human gene bank fall into not less than eight distinct classes, indicating that there are at least eight IFN-related genes. Most, if not all, of these direct the synthesis of an IFN in Escherichia coli. The sequence of one chromosomal gene and its flanking regions was identical to that deduced for the cDNA corresponding to IFN-αl mRNA. No evidence was found for the existence of an intron, in either the coding or the non-coding segments of the gene.
TL;DR: Recombinant plasmids capable of complementing cdc28 mutations were isolated by transformation of a cDC28ts strain and selection for clones capable of growth at the restrictive temperature, confirming the identity of the cloned sequences.
Abstract: cdc28, one of several genes required for cell division in the yeast Saccharomyces cerevisiae, has been isolated on recombinant plasmids. A recombinant plasmid pool containing the entire yeast genome was constructed by partial digestion of yeast DNA with the four-base recognition restriction endonuclease Sau3A to give the equivalent of random fragments, size selection on sucrose gradients, and introduction of the fragments into the yeast vector YRp7 by use of the homology of Sau3A ends with those generated in the vector by cleavage with BamHI. Recombinant plasmids capable of complementing cdc28 mutations were isolated by transformation of a cdc28ts strain and selection for clones capable of growth at the restrictive temperature. Plasmids responsible for complementing the cdc28ts phenotype were shown to recombine specifically with the chromosomal cdc28 locus, confirming the identity of the cloned sequences. In addition, one of the recombinant plasmids was capable of complementing a mutation in tyr1, a gene genetically linked to cdc28. This method of gene isolation and identification should be applicable to all yeast genes for which there are readily scorable mutants.
TL;DR: Circular double-stranded DNA produced after infection of mouse cells with Abelson murine leukemia virus was isolated and cloned in the phage vector Charon 21A and showed homology to the ends of Moloney MuLV and to a 3.5 kb central region containing sequences unique to Abelson virus.
TL;DR: The hamster gene coding for the enzyme adenine phosphoribosyl transferase (aprt) is isolated using gene transfer and molecular cloning of transforming DNA and sequences homologous to this clone are present in all hamster aprt+ transformants examined.
TL;DR: The DNA sequence of the wild type and mutated introns as well as their flanking exons in the yeast mitochondrial gene specifying cytochrome b, and a trans-acting protein "mRNA maturase" responsible for splicing and maturation of cy tochrome b mRNA are determined.
TL;DR: The results indicate that the leftmost 4.5% of Ads DNA is able to convert diploid cells in a primary culture into established aneuploid cell lines which still lack several of the properties characteristic for adenovirus 5-transformed cells.