2007 Fiscal Year Final Research Report Summary
Function analysis of mucin-type O-glycans on leukocytes in core 1 synthase knockout mouse.
| Project/Area Number |
17590236
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General medical chemistry
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| Research Institution | University of Tsukuba |
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Principal Investigator |
KUDO Takashi University of Tsukuba, Graduate School of Comprehensive Human Sciences, Associate Professor (20288062)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Satoru University of Tsukuba, Graduate School of Comprehensive Human Sciences, Professor (50271896)
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| Project Period (FY) |
2005 – 2007
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| Keywords | glycosyltransferasae / mucin / knockout mouse / galactosyltransferase |
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| Research Abstract |
The core 1 β1, 3-galactosyltransferase (T-synthase) transfers Gal from UDP-Gal to GalNAcα 1-Ser/Thr (Tn antigen) to form the core 1 O-glycan Gal β1-3 GalNAcalpha 1-Ser/Thr (T antigen). The T antigen is a precursor for extended and branched O-glycans of largely unknown function. We engineered conditional targeting vectors pDT-loxP-loxPFRT-PGKneo-loxPFRT, with three loxP sites and two FRT sites in the C1Gal-T or cosmc allele. The targeting vector were electrophorated into ES cells that were derived from a C57BL/6J mouse. ES cells clones with correct homologous recombination were selected by PCR and confirmed by Southern hybridization. Chimeras among the offspring were bred with Ayu-1 Cre or Flpe mice. Genotypes of mice were determined by PCR from tail biopsies. Matings between C1Gal-T+/-mice did not identified C1Gal-T knockout mice. Moreover, we established a polyclonal antibody against C1Gal-T polypeptides and investigated the localization of C1Gal-T protein in pancreas and stomach sections. C1Gal-T was present perinuclearly in endocrine cells of pancreas and secretory epithelial cells of stomach. We were identified that insulin 2-Cre/C1Gal-T^<flex/-> mice present compete and specific disruption of the C1Gal-T gene in pancreatic islet cells using immunohistochemical analysis. Mating of cosmc null mice are progressing now.
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