2000年3月
Cloning of the human cDNA which can complement the defect of the yeast mannosyltransferase I-deficient mutant alg 1
GLYCOBIOLOGY
- ,
- ,
- 巻
- 10
- 号
- 3
- 開始ページ
- 321
- 終了ページ
- 327
- 記述言語
- 英語
- 掲載種別
- DOI
- 10.1093/glycob/10.3.321
- 出版者・発行元
- OXFORD UNIV PRESS
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-P and that the function of this enzyme was conserved between yeast and human.
- リンク情報
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- DOI
- https://doi.org/10.1093/glycob/10.3.321
- CiNii Articles
- http://ci.nii.ac.jp/naid/80011753677
- PubMed
- https://www.ncbi.nlm.nih.gov/pubmed/10704531
- Web of Science
- https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000085571200011&DestApp=WOS_CPL
- ID情報
-
- DOI : 10.1093/glycob/10.3.321
- ISSN : 0959-6658
- CiNii Articles ID : 80011753677
- PubMed ID : 10704531
- Web of Science ID : WOS:000085571200011