Cloning the mouse homolog of the human cystic fibrosis transmembrane conductance regulator gene.
The cystic fibrosis transmembrane conductance regulator is encoded by the gene known to be mutated in patients with cystic fibrosis. This paper reports the cloning and sequencing of cDNAs for the murine homolog of the human cystic fibrosis transmembrane conductance regulator gene. A clone that, by analogy to the human sequence, extends 3' from exon 9 to the poly(A) tail was isolated from a mouse lung cDNA library. cDNA clones containing exons 4 and 6b were also isolated and sequenced, but the remainder of the mRNA proved difficult to obtain by conventional cDNA library screening. Sequences spanning exons 1-9 were cloned by PCR from mouse RNA. The deduced mouse protein sequence is 78% identical to the human cystic fibrosis transmembrane regulator, with higher conservation in the transmembrane and nucleotide-binding domains. Amino acid sequences in which known cystic fibrosis missense mutations occur are conserved between man and mouse; in particular, the predicted mouse protein has a phenylalanine residue corresponding to that deleted in the most common human cystic fibrosis mutation (delta F508), which should allow the use of transgenic strategies to introduce this mutation in attempts to create a "cystic fibrosis mouse".
Tata
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Department of Biochemistry and Molecular Genetics, St. Mary's Hospital Medical School, Imperial College, London, United Kingdom.
Stanier
P
P
Wicking
C
C
Halford
S
S
Kruyer
H
H
Lench
N J
NJ
Scambler
P J
PJ
Hansen
C
C
Braman
J C
JC
Williamson
R
R
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003550:Cystic Fibrosis / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA; D015723:Gene Library; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000138:chemical synthesis; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment
C505032:CFTR protein, human; D008565:Membrane Proteins; D009841:Oligonucleotides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA