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Cloning and expression of a porcine UDP-GalNAc: polypeptideN-acetylgalactosaminyl transferase

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Abstract

By employing a bovine UDP-N-acetylgalactosamine: polypeptideN-acetylgalactosaminyl transferase (O-GalNAc transferase) cDNA as a probe, we isolated four overlapping cDNAs from a porcine lung cDNA library. Both the nucleotide sequence of the porcine cDNA and the predicted primary structure of the protein (559 amino acids) proved to be very similar to those of the bovine enzyme (95% and 99% identity, respectively). Transient expression of the clone in COS-7 cells, followed by enzymatic activity assays, demonstrated that this cDNA sequence encodes a porcine O-GalNAc transferase. The intracellular O-GalNAc transferase activity was increased approximately 100-fold by transfecting cells with the porcine cDNA.

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Abbreviations

O-GalNAc transferase:

UDP-N-acetylgalactosamine: polypeptideN-acetylgalactosaminyltransferase

PCR:

polymerase chain reaction

SDS:

sodium dodecyl sulfate

PAGE:

polyacrylamide gel electrophoresis

GnT-III:

UDP-N-acetylglucosamine: β-mannoside β-1,4N-acetylglucosaminyltransferase III

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Authors and Affiliations

  1. Central Laboratories for Key Technology, Kirin Brewery Co., Ltd, Fukuura, Kanazawa-ku, 236, Yokohama, Japan

    Aruto Yoshida, Tomoka Hara, Hiroshi Ikenaga & Makoto Takeuchi

Authors
  1. Aruto Yoshida
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  2. Tomoka Hara
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  3. Hiroshi Ikenaga
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  4. Makoto Takeuchi
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Yoshida, A., Hara, T., Ikenaga, H. et al. Cloning and expression of a porcine UDP-GalNAc: polypeptideN-acetylgalactosaminyl transferase. Glycoconjugate J 12, 824–828 (1995). https://doi.org/10.1007/BF00731244

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  • DOI: https://doi.org/10.1007/BF00731244

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