Identification of protein coding regions by database similarity search

Gish, Warren; States, David J.

doi:10.1038/ng0393-266

Cited by 1,562 publications

(997 citation statements)

References 30 publications

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“…A TBLASTN search [20] was performed against the Expressed Sequence Tag (EST) database with the use of default parameters with reference to the predicted amino acid sequences of both the entire S. cere isiae Cpt1p-coding region and the Cpt1p CDPalcohol phosphotransferase motif [18]. Several ESTs were identified that possessed similarity to either the Cpt1p CDP-alcohol phosphotransferase motif or to other regions within the Cpt1p protein.…”

Section: Isolation and Expression Of The Full-length Hcept1mentioning

confidence: 99%

Cloning and expression of a human choline/ethanolaminephosphotransferase: synthesis of phosphatidylcholine and phosphatidylethanolamine

Henneberry

McMaster

1999

Biochemical Journal

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Cholinephosphotransferase catalyses the final step in the synthesis of phosphatidylcholine (PtdCho) via the Kennedy pathway by the transfer of phosphocholine from CDP-choline to diacylglycerol. Ethanolaminephosphotransferase catalyses an analogous reaction with CDP-ethanolamine as the phosphobase donor for the synthesis of phosphatidylethanolamine (PtdEtn). Together these two enzyme activities determine both the site of synthesis and the fatty acyl composition of PtdCho and PtdEtn synthesized de novo. A human choline/ethanolaminephosphotransferase cDNA (hCEPT1) was cloned, expressed and characterized. Northern blot analysis revealed one hCEPT1 2.3 kb transcript that was ubiquitous and not enriched, with respect to actin, in any particular cell type. The open reading frame predicts a protein (hCEPT1p) of 416 amino acid residues with a molecular mass of 46550 Da containing seven membrane-spanning domains. A predicted amphipathic helix resides within the active site of the enzyme with the final two aspartic residues of the CDP-alcohol phosphotransferase motif, DG(X)2AR(X)8G(X)3D(X)3D, positioned within this helix. hCEPT1p was successfully expressed in a full-length, active form in Saccharomyces cerevisiae cells devoid of endogenous cholinephosphotransferase or ethanolaminephosphotransferase activities (HJ091, cpt1::LEU2 ept1-). In vitro, hCEPT1p displayed broad substrate specificity, utilizing both CDP-choline and CDP-ethanolamine as phosphobase donors to a broad range of diacylglycerols, resulting in the synthesis of both PtdCho and PtdEtn. In vivo, S. cerevisiae cells (HJ091, cpt1::LEU2 ept1-) expressing hCEPT1 efficiently incorporated both radiolabelled choline and ethanolamine into phospholipids, demonstrating that hCEPT1p has the ability to synthesize both choline- and ethanolamine- containing phospholipids in vitro and in vivo.

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Section: Isolation and Expression Of The Full-length Hcept1mentioning

confidence: 99%

Cloning and expression of a human choline/ethanolaminephosphotransferase: synthesis of phosphatidylcholine and phosphatidylethanolamine

Henneberry

McMaster

1999

Biochemical Journal

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“…Nucleotide sequences in both directions of overlapping deletion clones were determined by the procedure of Sanger et al (1977), using 7-deaza dGTP to clarify regions previously obscured by GC compression (Mizusawa et al, 1986). Sequence data were compiled and comparative analyses were made using the GCG program (University of Wisconsin, Madison), the BLASTN, BLASTX, and BESTFIT programs developed by the National Centre for Biotechnology Information (Altschul et al, 1990;Devereux et al, 1984;Gish and States, 1993), and the GENECOMPAR program (Applied Maths Department, Kortrijk, Belgium).…”

Section: Dna Preparationmentioning

confidence: 99%

**Characterization of flagella genes of Agrobacterium tumefaciens, and the effect of a bald strain on virulence**

Chesnokova

Coutinho

Khan

et al. 1997

Molecular Microbiology

104

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Because the 54 promoter is associated with genes regulated by physiological changes in various bacteria, the flaC gene might be similarly regulated in response to A. tumefaciens responding to host plant stimuli. Virulence studies showed that the bald strain was consistently reduced in virulence below that of the parental wild-type strain by at least 38%. The difference is statistically significant and suggests that the flagella may play a role in facilitating virulence.

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“…1.0 (Cabot and Beckenbach 1989) and GCG (Genetic Computer Group, Madison, Wis.). Homology searches of the sequence databases were performed with the BLASTN and BLASTX algorithms (Altschul et al 1990;Gish and States 1993) using NCBI (National Center for Biotechnology Information) facilities accessed through the network server.…”

Section: Structure Of Psr and Pmentioning

confidence: 99%

Hybrid origin of a B chromosome (PSR) in the parasitic wasp Nasonia vitripennis

McAllister

Werren

1997

Chromosoma

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Abstract. Little is known about the origin and evolution of supernumerary (B) chromosomes. This study utilizes molecular markers to examine the evolutionary history and microstructural organization of the supernumerary paternal-sex-ratio (PSR) chromosome of the parasitic wasp Nasonia vitripennis. Copies of the retrotransposon NATE were previously isolated from PSR and the genomes of N. vitripennis and related wasp species. A phylogenetic analysis of sequences representing 29 elements from PSR and seven wasp species, coupled with a hybridization analysis of elements in genomic DNA provides evidence that PSR was recently transferred into N. vitripennis from a species in the genus Trichomalopsis. A linear region of the PSR chromosome was compared by Southern blot analysis with genomic DNA from N. vitripennis, Nasonia longicornis, Trichomalopsis americanus, and Trichomalopsis dubius. A region organized similarly to the region on PSR was not evident in any of the species, thus a progenitor region was not identified. However, the hybridizations revealed that this region of PSR is primarily composed of repetitive sequences that appear dispersed in these wasp genomes, and might represent additional mobile elements. At least three different dispersed repeats are present in the 18 kb region of PSR. The abundance of tandem and dispersed repetitive sequences in this relatively small region provides additional evidence for the degenerate structure of the PSR chromosome.

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Identification of protein coding regions by database similarity search

Cited by 1,562 publications

References 30 publications

Cloning and expression of a human choline/ethanolaminephosphotransferase: synthesis of phosphatidylcholine and phosphatidylethanolamine

Cloning and expression of a human choline/ethanolaminephosphotransferase: synthesis of phosphatidylcholine and phosphatidylethanolamine

**Characterization of flagella genes of Agrobacterium tumefaciens, and the effect of a bald strain on virulence**

Hybrid origin of a B chromosome (PSR) in the parasitic wasp Nasonia vitripennis

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