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.
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Research Article|
May 15 1995
Isolation and characterization of cDNA clones encoding pig gastric mucin
B S Turner;
B S Turner
*Section of Gastroenterology, Evans Department of Clinical Research, Section of Molecular Genetics, Boston University Medical Center, 88 East Newton Street, Boston, MA 02118, U.S.A.
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K R Bhaskar;
K R Bhaskar
*Section of Gastroenterology, Evans Department of Clinical Research, Section of Molecular Genetics, Boston University Medical Center, 88 East Newton Street, Boston, MA 02118, U.S.A.
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M Hadzopoulou-Cladaras;
M Hadzopoulou-Cladaras
†Department of Cellular Biology, School of Medicine in Shreveport, Louisiana State University Medical Center, 1501 Kings Highway, Shreveport, LA 71130, U.S.A.
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R D Specian;
R D Specian
†Department of Cellular Biology, School of Medicine in Shreveport, Louisiana State University Medical Center, 1501 Kings Highway, Shreveport, LA 71130, U.S.A.
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J T LaMont
J T LaMont
*Section of Gastroenterology, Evans Department of Clinical Research, Section of Molecular Genetics, Boston University Medical Center, 88 East Newton Street, Boston, MA 02118, U.S.A.
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Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1995 The Biochemical Society, London
1995
Biochem J (1995) 308 (1): 89–96.
Citation
B S Turner, K R Bhaskar, M Hadzopoulou-Cladaras, R D Specian, J T LaMont; Isolation and characterization of cDNA clones encoding pig gastric mucin. Biochem J 15 May 1995; 308 (1): 89–96. doi: https://doi.org/10.1042/bj3080089
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