Cloning of ClC-2 chloride channel from murine duodenum and its presence in CFTR knockout mice.
We report the cloning of a murine ClC-2 chloride channel cDNA from duodenal epithelium by reverse transcriptase-polymerase chain reaction (RT-PCR) using degenerate primers and by rapid amplification of cDNA ends (RACE)-PCR. Other than CFTR, this represents the first cloned chloride channel from intact intestine. The ClC-2 cDNA predicts encoding of a 908 amino acid polypeptide with a calculated M(r) of 99,373. The amino acid sequence of the murine ClC-2 chloride channel is over 94% identical to the ClC-2 chloride channel proteins of other species. Of interest is the finding that the ClC-2 mRNA is expressed about the same level in duodena from both CFTR knockout and wild-type mice. This is in keeping with the suggestion that ClC-2 might be a therapeutic target in cystic fibrosis.
Joo
N S
NS
Department of Pharmacology, University of Missouri, Columbia 65212, USA.
Clarke
L L
LL
Han
B H
BH
Forte
L R
LR
Kim
H D
HD
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D000090007:CLC-2 Chloride Channels; D018118:Chloride Channels / Q000235:genetics*; D003001:Cloning, Molecular; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D004386:Duodenum / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D010957:Plasmids; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment
D000090007:CLC-2 Chloride Channels; D018118:Chloride Channels; D018076:DNA, Complementary; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D012313:RNA