Tata F, Stanier P, Wicking C, Halford S, Kruyer H, Lench N J, Scambler P J, Hansen C, Braman J C, Williamson R
Department of Biochemistry and Molecular Genetics, St. Mary's Hospital Medical School, Imperial College, London, United Kingdom.
Genomics. 1991 Jun;10(2):301-7. doi: 10.1016/0888-7543(91)90312-3.
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".
囊性纤维化跨膜传导调节因子由已知在囊性纤维化患者中发生突变的基因编码。本文报道了人类囊性纤维化跨膜传导调节因子基因的小鼠同源物cDNA的克隆和测序。从小鼠肺cDNA文库中分离出一个克隆，根据与人类序列的类比，该克隆从外显子9延伸至3'端的多聚腺苷酸尾。还分离并测序了包含外显子4和6b的cDNA克隆，但通过常规cDNA文库筛选难以获得其余的mRNA。通过PCR从小鼠RNA中克隆了跨越外显子1 - 9的序列。推导的小鼠蛋白质序列与人类囊性纤维化跨膜调节因子有78%的同一性，在跨膜和核苷酸结合结构域中具有更高的保守性。已知囊性纤维化错义突变发生的氨基酸序列在人和小鼠之间是保守的；特别是，预测的小鼠蛋白质有一个苯丙氨酸残基，对应于最常见的人类囊性纤维化突变（ΔF508）中缺失的那个苯丙氨酸残基，这应该允许使用转基因策略引入该突变，试图创建一个“囊性纤维化小鼠”。
