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Cloning and Expression of Bovine Glucose Transporter GLUT12

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Abstract

GLUT12 is a new member of facilitative glucose transporters. It was originally cloned from a human breast cancer cell line and its expression has been detected in rat mammary gland. Glucose transport across the plasma membrane of mammary epithelial cells is a rate-limiting factor in milk production. To examine GLUT12’s expression and facilitate the study of GLUT12’s potential role in supporting milk synthesis in lactating bovine mammary gland, we cloned bovine GLUT12 and examined its distribution of mRNA expression in bovine tissues. The full-length mRNA of bGLUT12 is 2423 base pairs long and is predicted to encode a protein of 621 amino acids with a molecular weight of approximately 67 kDa. The deduced amino acid sequence of bovine GLUT12 is 87% and 82% identical to the sequences of human and mouse GLUT12. The sequence of bGLUT12 contains several characteristically conserved sugar transporter family signatures. Analysis of current bovine genomic data indicates that bovine GLUT12 gene consists of five exons. The major in vitro transcription and translation product of bovine GLUT12 cDNA migrated at an apparent molecular weight of 41 kDa. In the presence of canine microsomal membranes, the translation product increased to 43 kDa, suggesting glycosylation. GLUT12 mRNA was found in all bovine tissues examined, but most abundant in bovine spleen and skeletal muscle, at intermediate levels in bovine kidney, testes, and mammary gland, and at lower levels in bovine liver, lung and intestine. Immunofluorescence staining showed that, in the presence of insulin, bGLUT12 is mainly distributed in the cytoplasm of the transiently transfected MAC-T bovine mammary epithelial cells.

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Abbreviations

bGLUT12 = :

bovine solute carrier family 2 member 12 (gene symbol SLC2A12),

EST = :

expressed sequence tag,

kb = :

kilobases,

poly(A)+ = :

polyadenylated RNA,

RACE = :

rapid amplification of cDNA ends,

RT-PCR = :

reverse transcription-polymerase chain reaction,

TM = :

transmembrane domain,

UTR = :

untranslated region.

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Acknowledgments

This project was supported by National Research Initiative Competitive Grant No. 2005-35206-15267 from the USDA Cooperative State Research, Education, and Extension Service (to FQZ).

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Authors and Affiliations

  1. Lactation and Mammary Gland Biology Group, Department of Animal Science, University of Vermont, RM 219 Terrill Building,570 Main Street, Burlington, Vermont, 05405, USA

    Peter J. Miller, Kiera A. Finucane, Megan Hughes & Feng-Qi Zhao

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  1. Peter J. Miller
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  2. Kiera A. Finucane
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  3. Megan Hughes
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  4. Feng-Qi Zhao
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Corresponding author

Correspondence to Feng-Qi Zhao.

Additional information

The nucleotide sequence data reported in this paper have been submitted to GenBank with assigned accession number AY514443.

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Miller, P.J., Finucane, K.A., Hughes, M. et al. Cloning and Expression of Bovine Glucose Transporter GLUT12. Mamm Genome 16, 873–883 (2005). https://doi.org/10.1007/s00335-005-0080-5

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  • DOI: https://doi.org/10.1007/s00335-005-0080-5

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