Source:http://linkedlifedata.com/resource/pubmed/id/10704531
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
2000-3-17
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pubmed:abstractText |
The assembly of the lipid-linked oligosaccharide, Glc(3)Man(9)GlcNAc(2)-P-P-Dol, occurs on the rough ER membrane in an ordered stepwise manner. The process is highly conserved among eukaryotes. In order to isolate the human mannosyltransferase I (MT-I) gene involved in the process, we used the Saccharomyces cerevisiae MT-I gene ( ALG1 ), which has already been cloned. On searching the EST database with the amino acid sequence of the ALG1 gene product, we detected seven related human EST clones. A human fetal brain cDNA library was screened by PCR using gene-specific primers based on the EST nucleotide sequences and a 430 bp cDNA fragment was amplified. The cDNA library was rescreened with this 430 bp cDNA, and two cDNA clones (HR1-3 and HR1-4) were isolated and sequenced. On a homology search of the EST database with the nucleotide sequence of HR1-3, we detected a novel human EST clone, AA675921 (GenBank accession number). Based on the nucleotide sequences of AA675921 and HR1-4, we designed gene-specific PCR primers, which allowed to amplify a 1.8 kb cDNA from human fetal brain cDNA. This cDNA was cloned and shown to contain an ORF encoding a protein of 464 amino acids. We designated this ORF as Hmat-1. The amino acid sequence deduced from the Hmat-1 gene showed several highly conserved regions shared with the yeast and nematode MT-I sequences. Furthermore, this 1.8 kb cDNA successfully complemented the S. cerevisiae alg1-1 mutation, indicating that the Hmat-1 gene encodes the human MT-I and that the function of this enzyme was conserved between yeast and human.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
0959-6658
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
10
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
321-7
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:10704531-Amino Acid Sequence,
pubmed-meshheading:10704531-Base Sequence,
pubmed-meshheading:10704531-Cloning, Molecular,
pubmed-meshheading:10704531-DNA, Complementary,
pubmed-meshheading:10704531-Expressed Sequence Tags,
pubmed-meshheading:10704531-Genetic Complementation Test,
pubmed-meshheading:10704531-Humans,
pubmed-meshheading:10704531-Mannosyltransferases,
pubmed-meshheading:10704531-Molecular Sequence Data,
pubmed-meshheading:10704531-Mutation,
pubmed-meshheading:10704531-Saccharomyces cerevisiae,
pubmed-meshheading:10704531-Sequence Homology, Amino Acid
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pubmed:year |
2000
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pubmed:articleTitle |
Cloning of the human cDNA which can complement the defect of the yeast mannosyltransferase I-deficient mutant alg 1.
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pubmed:affiliation |
Laboratory for Glycobiology and Glycotechnology, Department of Industrial Chemistry, School of Engineering, Tokai University, 1117 Kitakaname, Hiratsuka City, Kanagawa 259-1292, Japan.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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