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Isolation and characterization of cDNA clones encoding pig gastric mucin.

Publication ,  Journal Article
Turner, BS; Bhaskar, KR; Hadzopoulou-Cladaras, M; Specian, RD; LaMont, JT
Published in: The Biochemical journal
May 1995

Polyclonal antibodies raised to deglycosylated pig gastric mucin were used to screen a cDNA library constructed with pig stomach mucosal mRNA. Immunocytochemistry indicated that the antibody recognizes intracellular and secreted mucin in surface mucous cells of pig gastric epithelium. A total of 70 clones producing proteins immunoreactive to this antibody were identified, two of which (PGM-2A,9B) were fully sequenced from both ends. Clone PGM-9B hybridized to a polydisperse mRNA (3-9 kb) from pig stomach, but not liver, intestine or spleen, nor to mRNA from human, mouse, rabbit or rat stomach. Sequence analysis indicated that PGM-9B encodes 33 tandem repeats of a 16-amino-acid consensus sequence rich in serine (46%) and threonine (17%). Using the restriction enzyme MwoI, which has a single target site in the repeat, it was demonstrated that PGM-9B consists entirely of this tandem repeat. Southern-blot analysis indicated that the repeat region is contained in a 20 kb HindIII-EcoRI fragment, and BamHI digestion suggested that most of the repeats are contained in a 10 kb fragment. In situ hybridization with an antisense probe to PGM-9B showed an intense signal in the entire gastric gland. Clone PGM-2A also contains the same repeat sequence as 9B, but, in addition, has a 64-amino-acid-long non-repeat region at its 5' end. Interestingly the non-repeat region of PGM-2A has five cysteine residues, the arrangement of which is identical with that reported for human intestinal mucin gene MUC2.

Duke Scholars

Published In

The Biochemical journal

DOI

EISSN

1470-8728

ISSN

0264-6021

Publication Date

May 1995

Volume

308 ( Pt 1)

Start / End Page

89 / 96

Related Subject Headings

  • Swine
  • Sequence Alignment
  • Repetitive Sequences, Nucleic Acid
  • RNA, Messenger
  • Mucins
  • Molecular Sequence Data
  • In Situ Hybridization
  • Genes
  • Gene Expression
  • Gastric Mucosa
 

Citation

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Turner, B. S., Bhaskar, K. R., Hadzopoulou-Cladaras, M., Specian, R. D., & LaMont, J. T. (1995). Isolation and characterization of cDNA clones encoding pig gastric mucin. The Biochemical Journal, 308 ( Pt 1), 89–96. https://doi.org/10.1042/bj3080089
Turner, B. S., K. R. Bhaskar, M. Hadzopoulou-Cladaras, R. D. Specian, and J. T. LaMont. “Isolation and characterization of cDNA clones encoding pig gastric mucin.The Biochemical Journal 308 ( Pt 1) (May 1995): 89–96. https://doi.org/10.1042/bj3080089.
Turner BS, Bhaskar KR, Hadzopoulou-Cladaras M, Specian RD, LaMont JT. Isolation and characterization of cDNA clones encoding pig gastric mucin. The Biochemical journal. 1995 May;308 ( Pt 1):89–96.
Turner, B. S., et al. “Isolation and characterization of cDNA clones encoding pig gastric mucin.The Biochemical Journal, vol. 308 ( Pt 1), May 1995, pp. 89–96. Epmc, doi:10.1042/bj3080089.
Turner BS, Bhaskar KR, Hadzopoulou-Cladaras M, Specian RD, LaMont JT. Isolation and characterization of cDNA clones encoding pig gastric mucin. The Biochemical journal. 1995 May;308 ( Pt 1):89–96.

Published In

The Biochemical journal

DOI

EISSN

1470-8728

ISSN

0264-6021

Publication Date

May 1995

Volume

308 ( Pt 1)

Start / End Page

89 / 96

Related Subject Headings

  • Swine
  • Sequence Alignment
  • Repetitive Sequences, Nucleic Acid
  • RNA, Messenger
  • Mucins
  • Molecular Sequence Data
  • In Situ Hybridization
  • Genes
  • Gene Expression
  • Gastric Mucosa