Cloning and chromosomal mapping of the mouse Mgat3 gene encoding N-acetylglucosaminyltransferase III

Mantu Bhaumik, Michael F Seldin, Pamela Stanley

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Complex and hybrid N-linked carbohydrates synthesized by mammalian cells may possess a N-acetylglucosamine residue known as the bisecting GlcNAc. The transfer of this residue is catalyzed by the enzyme UDP-N-acetylglucosamine:β-d-mannoside β1,4-N-acetylglucosaminyltransferase III (GlcNAc-TIII; EC 2.4.1.144). To begin to investigate biological functions for carbohydrates with a bisected GlcNAc residue, we have cloned and partially characterized the mouse gene (Mgat3) encoding GlcNAc-TIII. A rat GlcNAc-TIII-encoding cDNA was used to isolate clones from a mouse strain 129 Sv liver genomic DNA library. An NsiI genomic DNA fragment containing an ORF with 96% identity to rat GlcNAc-TIII was subcloned into a mammalian expression vector and transfected into Chinese hamster ovary (CHO) cells. The transfectants expressed GlcNAc-TIII activity only when the ORF was in the sense orientation. Southern analysis showed that Mgat3 is present in a single copy in the mouse genome. Mapping by restriction-fragment length polymorphism analysis of backcross progeny located Mgat3 to mouse chromosome 15, at a position homologous with region 22q12.3-q13.1 in the human genome. Northern analyses of adult tissues showed that Mgat3 is expressed at high levels in kidney and brain, and at lower levels in many other tissues.

Original languageEnglish (US)
Pages (from-to)295-300
Number of pages6
JournalGene
Volume164
Issue number2
DOIs
StatePublished - Oct 27 1995
Externally publishedYes

Fingerprint

beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase
Organism Cloning
Open Reading Frames
Carbohydrates
129 Strain Mouse
Uridine Diphosphate N-Acetylglucosamine
Mannosides
Genes
Chromosomes, Human, Pair 15
Acetylglucosamine
Genomic Library
Human Genome
Cricetulus
Gene Library
Restriction Fragment Length Polymorphisms
Ovary
Complementary DNA
Clone Cells
Genome
Kidney

Keywords

  • mouse genome
  • Northern analysis
  • transfection

ASJC Scopus subject areas

  • Genetics

Cite this

Cloning and chromosomal mapping of the mouse Mgat3 gene encoding N-acetylglucosaminyltransferase III. / Bhaumik, Mantu; Seldin, Michael F; Stanley, Pamela.

In: Gene, Vol. 164, No. 2, 27.10.1995, p. 295-300.

Research output: Contribution to journalArticle

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abstract = "Complex and hybrid N-linked carbohydrates synthesized by mammalian cells may possess a N-acetylglucosamine residue known as the bisecting GlcNAc. The transfer of this residue is catalyzed by the enzyme UDP-N-acetylglucosamine:β-d-mannoside β1,4-N-acetylglucosaminyltransferase III (GlcNAc-TIII; EC 2.4.1.144). To begin to investigate biological functions for carbohydrates with a bisected GlcNAc residue, we have cloned and partially characterized the mouse gene (Mgat3) encoding GlcNAc-TIII. A rat GlcNAc-TIII-encoding cDNA was used to isolate clones from a mouse strain 129 Sv liver genomic DNA library. An NsiI genomic DNA fragment containing an ORF with 96{\%} identity to rat GlcNAc-TIII was subcloned into a mammalian expression vector and transfected into Chinese hamster ovary (CHO) cells. The transfectants expressed GlcNAc-TIII activity only when the ORF was in the sense orientation. Southern analysis showed that Mgat3 is present in a single copy in the mouse genome. Mapping by restriction-fragment length polymorphism analysis of backcross progeny located Mgat3 to mouse chromosome 15, at a position homologous with region 22q12.3-q13.1 in the human genome. Northern analyses of adult tissues showed that Mgat3 is expressed at high levels in kidney and brain, and at lower levels in many other tissues.",
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