Isolation and characterization of cDNA clones encoding pig gastric mucin.
Journal Article (Journal Article)
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.
Full Text
Duke Authors
Cited Authors
- Turner, BS; Bhaskar, KR; Hadzopoulou-Cladaras, M; Specian, RD; LaMont, JT
Published Date
- May 1995
Published In
Volume / Issue
- 308 ( Pt 1) /
Start / End Page
- 89 - 96
PubMed ID
- 7755593
Pubmed Central ID
- PMC1136847
Electronic International Standard Serial Number (EISSN)
- 1470-8728
International Standard Serial Number (ISSN)
- 0264-6021
Digital Object Identifier (DOI)
- 10.1042/bj3080089
Language
- eng