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C Helms

Publications and source records attributed to C Helms.

33 records · Page 2Linked to original sources
PubMed1989

Identification and characterization of 23 RFLP loci by screening random cosmid genomic clones.

As part of our search for polymorphic DNA probes, we have screened cosmids from a human genomic DNA library for their ability to reveal RFLPs. A total of 101 randomly isolated cosmid clones were tested in Southern hybridizations for polymorphic band patterns. Fifty-four of these clones revealed RFLPs with one or more of nine restriction enzymes. Twenty-three of these clones have been further characterized and assigned to 10 different chromosomes by linkage analysis or by hybridization to panels of human-hamster hybrid cell lines. Fifteen of the probes have heterozygosities greater than or equal to .5. The relative efficiency of RsaI and PstI restriction enzymes in detecting polymorphism was different from results obtained with libraries constructed in bacteriophage vectors. Screening randomly selected cosmid probes is an efficient method for detecting RFLPs.

Alleles↗
PubMed1989

Construction of a linkage map of the human genome, and its application to mapping genetic diseases.

A powerful method for localizing the genes responsible for genetic diseases is to "mark" each chromosomal region so as to permit its inheritance to be tracked in families carrying the disease; the disease gene must lie in the same region as any marker with which it is co-inherited. Chromosomes can be efficiently marked in this way by using restriction fragment length polymorphisms (RFLPs). We describe the construction of a map of the human genome with over 550 RFLP markers, such that over 95% of the genome is detectably linked to one or more of these markers. This map will make possible efficient searches for genes underlying a variety of disorders, both simple and complex, and lead to the development of presymptomatic diagnostic tests for these diseases.

Base Sequence↗
PubMed1988

Automated percutaneous discectomy: preliminary experience.

In 1985, Onik, described a new automated lumbar disc aspiration technique that utilizes a probe with the guillotine cutting technology used in vitrectomy and arthroscopic instrumentation. The probe's small size (2 mm in diameter) minimizes the risk of nerve injury, while its automated action permits rapid, safe removal of disc material. The technique and our preliminary results in patients treated with this method are described.

Dermatologic Surgical Procedures↗
PubMed1988

Beta 2-adrenergic mechanisms in experimental arthritis.

We have studied (i) the contribution of specific adrenergic receptors to the proinflammatory effects of the sympathetic nervous system in experimental arthritis and (ii) the phases of the disease during which the sympathetic nervous system influences joint injury. Severity of joint injury was measured radiographically 28 days after induction of adjuvant arthritis in control rats and in rats treated with a variety of sympatholytic agents at various times during the course of the disease. Rats treated with a nonspecific catecholamine depletor (reserpine) or a beta-adrenergic receptor antagonist (propranolol) had a delayed onset and significantly less severe joint injury than saline-treated controls when treatment began prior to injection of the adjuvant and continued to day 28 after the injection. When administered over the same treatment period, neither nonselective (phenoxybenzamine) nor selective [prazosin (alpha 1) and yohimbine (alpha 2)] alpha-adrenergic receptor antagonists affected the onset or severity of joint injury. Metoprolol, a beta 1 antagonist, was also without effect. In contrast, two beta 2 antagonists (butoxamine and ICI 118,551) significantly retarded disease onset and reduced the severity of joint injury. When reserpine or butoxamine treatment was initiated after the onset of clinically apparent arthritis, it was still possible to favorably influence the course of the disease. These data indicate an important contribution of the beta 2-adrenergic receptor to joint injury in experimental arthritis.

Animals↗
PubMed1988

Identification of more than 500 RFLPs by screening random genomic clones.

As part of our genome-mapping effort, we undertook a large-scale screening study to identify RFLPs useful as genetic markers. Some 1,664 single-copy or repeat-containing phage clones from a Charon 4A genomic library were tested for polymorphism against a panel of DNAs, from five unrelated individuals, digested with eight restriction enzymes. Approximately 30% (515) of the clones revealed polymorphism by Southern hybridization; 67 loci detected had PIC values greater than .5. Restriction enzymes MspI, TaqI, and RsaI were most efficient in detecting polymorphism within the 1-20-kb-fragment size range resolved. With only one exception each of the clones detected polymorphism originating from a single locus.

Cloning, Molecular↗
PubMed1987

Concerted deletions and inversions are caused by mitotic recombination between delta sequences in Saccharomyces cerevisiae.

Deletions of a tyrosine tRNA suppressor gene, SUP4-o, are mediated by recombination between short repeated delta sequences in Saccharomyces cerevisiae. The arrangement of the five solo delta sequences that surround the SUP4 locus was established by DNA sequence analysis. Seven deletion classes were identified by genomic blotting. DNA sequence analysis also showed that the delta sequences within a 6.5-kilobase region of the SUP4 locus were the endpoints of these events. In three of these classes, an adjacent interval surrounded by delta sequences was inverted in concert with the deletion. The frequency of all deletion classes decreased in strains that contained mutations in the recombination and repair gene RAD52. We present two gene conversion mechanisms by which these rearrangements could have been generated. These models may also explain deletions between repeated sequences in other systems.

Base Sequence↗
PubMed1986

Factors which equalize the representation of genome segments in recombinant libraries.

Genomic segments which contain inverted repetitions longer than 300 bp are frequently lost from recombinant libraries grown on rec+ hosts. We have found that 9% of phage lambda clones that contain 15-20-kb insertions of human or Drosophila DNA are inhibited on rec+ hosts and as a result will become under-represented in amplified genomic libraries. We have therefore examined several factors of both host and vector origin which affect the fidelity of representation of genomic sequences in recombinant DNA libraries constructed in bacteriophage lambda vectors. This loss may be diminished if the vector carries either a chi element or a functional gam gene. The most successful approach, however, involves using a host with mutations in recB, recC, and sbcB, or in recD. We have shown that recombinant clones which require such mutant hosts for growth are somewhat more likely to contain DNA derived from loci in the genome which are polymorphic than are clones recovered on conventional hosts.

Animals↗
PubMed1986

Random-clone strategy for genomic restriction mapping in yeast.

An approach to global restriction mapping is described that is applicable to any complex source DNA. By analyzing a single restriction digest for each member of a redundant set of lambda clones, a data base is constructed that contains fragment-size lists for all the clones. The clones are then grouped into subsets, each member of which is related to at least one other member by a significant overlap. Finally, a tree-searching algorithm seeks restriction maps that are consistent with the fragment-size lists for all the clones in each subset. The feasibility of the approach has been demonstrated by collecting data on 5000 lambda clones containing random 15-kilobase inserts of yeast DNA. It is shown that these data can be analyzed to produce regional maps of the yeast genome, extending in some cases for over 100 kilobases. In combination with hybridization probes to previously cloned genes, these local maps are already useful for defining the physical arrangement of closely linked genes. They may in the future serve as building blocks for the construction of a continuous global map.

Chromosome Mapping↗
PubMed1985

A new method for purifying lambda DNA from phage lysates.

A new method for preparing small quantities of lambda DNA from phage lysates has been developed. The protocol is based on the concentration and purification of bacteriophage particles from crude lysates using small DEAE-cellulose columns. This chromatographic step gives an absolute separation of the lambda DNA from the cellular nucleic acids and a 20-fold enrichment relative to the major soluble proteins in crude lysates, while effecting a 10-fold concentration of the phage. Final deproteinization and concentration of the lambda DNA is achieved by conventional precipitation steps. The lambda DNA produced by this method is shown to be nondegraded, biologically active, and an excellent substrate for restriction enzymes. A detailed protocol is provided for starting with individual plaques and using the method to obtain purified DNA from large numbers of lambda clones.

Bacteriophage lambda↗
PubMed1978

Chromosome mapping of the CYC7 gene determining yeast iso-2-cytochrome c: structural and regulatory regions.

The primary structures of iso-1-cytochrome c and iso-2-cytochrome c in the yeast Saccharomyces cerevisiae are determined by the genes CYC1 and CYC7, respectively. The CYC1 locus was previously shown to be on the right arm of chromosome X, and the CYC7 locus is shown in this investigation to be on the left arm of chromosome V closely linked to the min1 and mak10 markers. The CYC7 locus appears to be composed of a structural region and a regulatory region. Mutations in the structural region can cause a deficiency or alteration of iso-2-cytochrome c, whereas mutations in the regulatory region can cause increases in the amount of iso-2-cytochrome c. Single-site gene conversion, occurring at a relatively high frequency of approximately 4%, caused intragenic recombination of a mutational site in the structural region and a mutational site in the regulatory region, enabling us to suggest the order of the sites in relationship to other markers on the chromosome.

Chromosome Mapping↗
PubMed1978

A chromosomal translocation causing overproduction of iso-2-cytochrome c in yeast.

The CYC7-1 mutation in the yeast Saccharomyces cerevisiae causes the production of approximately 30 times the normal amount of iso-2-cytochrome c. Genetic analysis established that the CYC7-1 mutation is a reciprocal translocation involving the left arm of chromosome V and the right arm of chromosome XVI. The chromosome V arm was broken adjacent to the gene CYC7, which determines the primary structure of iso-2-cytochrome c, and this fragment containing the CYC7 gene was joined to the segment of chromosome XVI. It appears as though the elevation of iso-2-cytochrome c is caused by an abnormal controlling region adjacent to the structural region of the CYC7 gene.

Cytochrome c Group↗
PubMedDate not supplied

The application of molecular and immunologic techniques to study the epidemiology of Legionella pneumophila serogroup 1.

We applied monoclonal antibody typing and restriction endonuclease analysis of plasmid DNA to study 28 clinical and 35 environmental (potable water) isolates of Legionella pneumophila serogroup 1 from three hospitals in Iowa between 1981 and 1986. Monoclonal antibody typing employed a panel of seven antibodies and delineated eight different subtypes. Plasmids were present in 57% of the isolates including 12 of 28 (43%) clinical and 25 of 35 (69%) potable water isolates. The plasmids ranged in size from 28 to 98 kilobase pairs and comprised eight distinct subtypes by restriction endonuclease analysis with Eco RI. Combination of monoclonal antibody and restriction endonuclease subtyping (composite subtyping) revealed 19 different composite subtypes of Legionella pneumophila serogroup 1. The most common composite subtype, 09:04, comprised 29% (18 of 63) of the isolates and was only found in clinical and potable water samples from a single pavilion in hospital A during an outbreak of Legionella pneumophila serogroup 1 pneumonia. Aside from this cluster the diversity of composite subtypes of Legionella pneumophila serogroup 1 observed in clinical and potable water sources over the 5-year period was striking. The combination of monoclonal antibody and restriction endonuclease typing resulted in improved strain delineation and a more useful use of epidemiologic markers for Legionella pneumophila serogroup 1.

Antibodies, Monoclonal↗