A highly polymorphic locus on chromosome 16q revealed by a probe from a chromosome-specific cosmid library

AbstractThe SSD1 gene of Saccharomyces cerevisiae is a polymorphic locus that affects diverse cellular processes including cell integrity, cell cycle progression, and growth at high temperature. We show here that the SSD1-V allele is necessary for cells to achieve extremely long life span. Furthermore, addition of SSD1-V to cells can increase longevity independently of SIR2, although SIR2 is necessary for SSD1-V cells to attain maximal life span. Past studies of yeast aging have been performed in short-lived ssd1-d strain backgrounds. We propose that SSD1-V defines a previously undescribed pathway affecting cellular longevity and suggest that future studies on longevity-promoting genes should be carried out in long-lived SSD1-V strains.

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A Susceptibility Gene for Psoriatic Arthritis Maps to Chromosome 16q: Evidence for Imprinting

The American Journal of Human Genetics ◽

10.1086/345646 ◽

2003 ◽

Vol 72 (1) ◽

pp. 125-131 ◽

Cited By ~ 121

Author(s):

Ari Karason ◽

Johann E. Gudjonsson ◽

Ruchi Upmanyu ◽

Arna A. Antonsdottir ◽

Valdimar B. Hauksson ◽

...

Keyword(s):

Psoriatic Arthritis ◽

Susceptibility Gene ◽

Chromosome 16Q

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Identification of Resistance Determinants for a Promising Antileishmanial Oxaborole Series

Microorganisms ◽

10.3390/microorganisms9071408 ◽

2021 ◽

Vol 9 (7) ◽

pp. 1408

Author(s):

Magali Van den Kerkhof ◽

Philippe Leprohon ◽

Dorien Mabille ◽

Sarah Hendrickx ◽

Lindsay B. Tulloch ◽

...

Keyword(s):

Drug Exposure ◽

In Vitro Selection ◽

Selection Procedure ◽

Cosmid Library ◽

Neglected Diseases ◽

Chromosome 2 ◽

Resistance Determinants ◽

A Genome

Current treatment options for visceral leishmaniasis have several drawbacks, and clinicians are confronted with an increasing number of treatment failures. To overcome this, the Drugs for Neglected Diseases initiative (DNDi) has invested in the development of novel antileishmanial leads, including a very promising class of oxaboroles. The mode of action/resistance of this series to Leishmania is still unknown and may be important for its further development and implementation. Repeated in vivo drug exposure and an in vitro selection procedure on both extracellular promastigote and intracellular amastigote stages were both unable to select for resistance. The use of specific inhibitors for ABC-transporters could not demonstrate the putative involvement of efflux pumps. Selection experiments and inhibitor studies, therefore, suggest that resistance to oxaboroles may not emerge readily in the field. The selection of a genome-wide cosmid library coupled to next-generation sequencing (Cos-seq) was used to identify resistance determinants and putative targets. This resulted in the identification of a highly enriched cosmid, harboring genes of chromosome 2 that confer a subtly increased resistance to the oxaboroles tested. Moderately enriched cosmids encompassing a region of chromosome 34 contained the cleavage and polyadenylation specificity factor (cpsf) gene, encoding the molecular target of several related benzoxaboroles in other organisms.

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Gene Cloning and Molecular Characterization of a Two-Enzyme System Catalyzing the Oxidative Detoxification of β-Endosulfan

Applied and Environmental Microbiology ◽

10.1128/aem.68.12.6237-6245.2002 ◽

2002 ◽

Vol 68 (12) ◽

pp. 6237-6245 ◽

Cited By ~ 36

Author(s):

Tara D. Sutherland ◽

Irene Horne ◽

Robyn J. Russell ◽

John G. Oakeshott

Keyword(s):

Shuttle Vector ◽

Open Reading Frame ◽

Cosmid Library ◽

Flavin Mononucleotide ◽

Flavin Reductase ◽

Reading Frame ◽

Reductase Gene ◽

Degradation Activity ◽

Layer Chromatography

ABSTRACT The gram-positive bacterium Mycobacterium sp. strain ESD is able to use the cyclodiene insecticide endosulfan as a source of sulfur for growth. This activity is dependent on the absence of sulfite or sulfate in the growth medium. A cosmid library of strain ESD DNA was constructed in a Mycobacterium-Escherichia coli shuttle vector and screened for endosulfan-degrading activity in Mycobacterium smegmatis, a species that does not degrade endosulfan. Using this method, we identified a single cosmid that conferred sulfur-dependent endosulfan-degrading activity on the host strain. An open reading frame (esd) was identified within this cosmid that, when expressed behind a constitutive promoter in a mycobacterial expression vector, conferred sulfite- and sulfate-independent β-endosulfan degradation activity on the recombinant strain. The translation product of this gene (Esd) had up to 50% sequence identity with an unusual family of monooxygenase enzymes that use reduced flavins, provided by a separate flavin reductase enzyme, as cosubstrates. An additional partial open reading frame was located upstream of the Esd gene that had sequence homology to the same monooxygenase family. A flavin reductase gene, identified in the M. smegmatis genome, was cloned, expressed, and used to provide reduced flavin mononucleotide for Esd in enzyme assays. Thin-layer chromatography and gas chromatography analyses of the enzyme assay mixtures revealed the disappearance of β-endosulfan and the appearance of the endosulfan metabolites, endosulfan monoaldehyde and endosulfan hydroxyether. This suggests that Esd catalyzes the oxygenation of β-endosulfan to endosulfan monoaldehyde and endosulfan hydroxyether. Esd did not degrade either α-endosulfan or the metabolite of endosulfan, endosulfan sulfate.

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Isolation of a functional human interleukin 2 gene from a cosmid library by recombination in vivo

Gene ◽

10.1016/0378-1119(85)90104-0 ◽

1985 ◽

Vol 39 (1) ◽

pp. 33-39 ◽

Cited By ~ 9

Author(s):

W. Lindenmaier ◽

K.E.J. Dittmar ◽

H. Hauser ◽

A. Necker ◽

W. Sebald

Keyword(s):

Interleukin 2 ◽

Cosmid Library

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A Type II Protein Secretory Pathway Required for Levansucrase Secretion by Gluconacetobacter diazotrophicus

Journal of Bacteriology ◽

10.1128/jb.186.15.5031-5039.2004 ◽

2004 ◽

Vol 186 (15) ◽

pp. 5031-5039 ◽

Cited By ~ 26

Author(s):

Juan G. Arrieta ◽

Mailin Sotolongo ◽

Carmen Menéndez ◽

Dubiel Alfonso ◽

Luis E. Trujillo ◽

...

Keyword(s):

Codon Usage ◽

Outer Membrane ◽

Gene Cluster ◽

Secretory Pathway ◽

Cosmid Library ◽

Polymorphism Analysis ◽

Type Ii ◽

Gluconacetobacter Diazotrophicus ◽

Type Iv ◽

Geographical Regions

ABSTRACT The endophytic diazotroph Gluconacetobacter diazotrophicus secretes a constitutively expressed levansucrase (LsdA, EC 2.4.1.10) to utilize plant sucrose. LsdA, unlike other extracellular levansucrases from gram-negative bacteria, is transported to the periplasm by a signal-peptide-dependent pathway. We identified an unusually organized gene cluster encoding at least the components LsdG, -O, -E, -F, -H, -I, -J, -L, -M, -N, and -D of a type II secretory system required for LsdA translocation across the outer membrane. Another open reading frame, designated lsdX, is located between the operon promoter and lsdG, but it was not identified in BLASTX searches of the DDBJ/EMBL/GenBank databases. The lsdX, -G, and -O genes were isolated from a cosmid library of strain SRT4 by complementation of an ethyl methanesulfonate mutant unable to transport LsdA across the outer membrane. The downstream genes lsdE, -F, -H, -I, -J, -L, -M, -N, and -D were isolated through chromosomal walking. The high G+C content (64 to 74%) and the codon usage of the genes identified are consistent with the G+C content and codon usage of the standard G. diazotrophicus structural gene. Sequence analysis of the gene cluster indicated that a polycistronic transcript is synthesized. Targeted disruption of lsdG, lsdO, or lsdF blocked LsdA secretion, and the bacterium failed to grow on sucrose. Replacement of Cys162 by Gly at the C terminus of the pseudopilin LsdG abolished the protein functionality, suggesting that there is a relationship with type IV pilins. Restriction fragment length polymorphism analysis revealed conservation of the type II secretion operon downstream of the levansucrase-levanase (lsdA-lsdB) locus in 14 G. diazotrophicus strains representing 11 genotypes recovered from four different host plants in diverse geographical regions. To our knowledge, this is the first report of a type II pathway for protein secretion in the Acetobacteraceae.

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A confirmed association between an ER-ß polymorphic locus and refuted associations in TIMP-1 and MMP-9 with AAA

British Journal of Surgery ◽

10.1002/bjs.6539 ◽

2009 ◽

Vol 96 (S1) ◽

pp. 13-13

Author(s):

H. S. Rayt ◽

M. J. Bown ◽

N. J. London ◽

R. D. Sayers

Keyword(s):

Polymorphic Locus

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A New Multilocus Probe for DNA Fingerprinting in Chinook Salmon (Oncorhynchus tshawytscha), and Comparisons with a Single-Locus Probe

Canadian Journal of Fisheries and Aquatic Sciences ◽

10.1139/f93-177 ◽

1993 ◽

Vol 50 (7) ◽

pp. 1559-1567 ◽

Cited By ~ 22

Author(s):

T. A. Stevens ◽

R. E. Withler ◽

S. H. Goh ◽

T. D. Beacham

Keyword(s):

Chinook Salmon ◽

Oncorhynchus Tshawytscha ◽

Polymorphic Locus ◽

Pedigree Analysis ◽

Single Locus ◽

Dna Fingerprint ◽

Dna Fragments ◽

Banding Patterns ◽

Discriminatory Ability ◽

Juvenile Chinook Salmon

A multilocus DNA probe, B2-2, isolated from chinook salmon (Oncorhynchus tshawytscha) and a single-locus Atlantic salmon (Salmo salar) probe, 3.15.34, were examined for discriminatory ability among seven parents and 33–37 juveniles from five families of chinook salmon. DNA fingerprint patterns were observed in Hae III-digested chinook salmon DNA probed with B2-2. Between 8 and 20 fragments, from 2.20 kilobase pairs (kbp) to 19.0 kbp, were detected in each individual. The level of band sharing among unrelated parents was 0.18. Probe 3.15.34 hybridized with a total of nine DNA fragments, from 3.35 to 6.00 kbp, in the chinook salmon parents and progeny. One or two fragments were detected in each individual. Pedigree analysis confirmed that 3.15.34 detected both alleles of a single polymorphic locus whereas B2-2 detected autosomal, unlinked, predominantly heterozygous DNA fragments that were inherited in a Mendelian fashion at a minimum of 10 polymorphic loci. Among juvenile chinook salmon, levels of band sharing detected with probe B2-2 increased with increasing relatedness, and clustering based on differences in banding patterns distinguished unrelated progeny, half sibs, and full sibs even in the absence of parental genotypic data.

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