Molecular cloning of rat intestinal mucin. Lack of conservation between mammalian species.
We have prepared antisera to deglycosylated rat intestinal mucin and used it to obtain immunoreactive clones from a rat jejunum cDNA library. Four of these clones were sequenced, and all were found to be partial cDNAs that contained 18-base pair tandem repeats characteristic of a mucin. These cDNAs encoded a repetitive peptide with a consensus sequence of TTTPDV. Thus, they bear little resemblance to either of the two human intestinal mucin cDNAs isolated previously (Gum, J. R., Byrd, J. C., Hicks, J. W., Toribara, N. W., Lamport, D. T. A., and Kim, Y. S. (1989) J. Biol. Chem. 264, 6480-6487 and Gum, J. R., Hicks, J. W., Swallow, D. M., Lagace, R. E., Byrd, J. C., Lamport, D. T. A., Siddiki, B., and Kim, Y. S. (1990) Biochem. Biophys. Res. Commun. 171, 407-415). One of these rat mucin clones, designated RMUC 176, was chosen for further analysis. This clone recognized a band of approximately 9 kilobases when used to probe RNA blots. A strong hybridization band was present using rat small intestine and colon RNA but was not detectable when RNA isolated from heart, liver, or kidney was tested. The RMUC 176 clone and the two previously isolated human intestinal mucin cDNA clones were used to probe blots prepared from BamHI-digested DNA of various species. Here, the human probes detected fragments present only in human and chimpanzee DNA, whereas the RMUC 176 clone recognized fragments only in rat and mouse DNA. Thus, the repetitive portions of intestinal mucin genes are apparently not well conserved between phylogenetically distant species.
Gastrointestinal Research Laboratory, Department of Veterans Affairs Medical Center, San Francisco, California 94121.
J R
JR
Gum
J W
JW
Hicks
R E
RE
Lagace
J C
JC
Byrd
N W
NW
Toribara
B
B
Siddiki
F J
FJ
Fearney
D T
DT
Lamport
Y S
YS
Kim
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D002241:Carbohydrates / Q000032:analysis; D002846:Chromatography, Affinity; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D015723:Gene Library; D007421:Intestine, Small / Q000502:physiology*; D007583:Jejunum / Q000502:physiology*; D008297:Male; D008969:Molecular Sequence Data; D009077:Mucins / Q000235:genetics* / Q000302:isolation & purification; D051381:Rats; D011919:Rats, Inbred Strains; D012091:Repetitive Sequences, Nucleic Acid
D000596:Amino Acids; D002241:Carbohydrates; D009077:Mucins; D004247:DNA