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Volumn 245, Issue 4922, 1989, Pages 1066-1073

Identification of the cystic fibrosis gene: Cloning and characterization of complementary DNA

Author keywords

[No Author keywords available]

Indexed keywords

NUCLEOTIDE BINDING PROTEIN;

EID: 0024424270     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.2475911     Document Type: Article
Times cited : (6047)

References (80)
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    • and used as template for the synthesis of cDNA according to After methylation of internal Eco RI sites, ends were made flush with T4 DNA polymerase, and phosphorylated Eco RI linkers were added to the cDNA. After digestion with Eco RI and removal of excess linkers, the cDNA products were ligated into the Eco RI site of λ ZAP (Stratagene, San Diego, CA)
    • (1983) Gene , vol.25 , pp. 263
    • Gubler, U.1    Hoffman, B.2
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    • 6 from T84 cells. Standard procedures were used for screening Cold Spring Harbor Laboratory, Cold Spring Harbor, NY Bluescript plasmids were rescued from plaque-purified clones by excision with M13 helper phage (Stratagene). The lung and pancreas libraries were purchased from Clontech Lab Inc. (Catalog Nos. HL1066b and HL1069h, respectively)
    • (1982) Molecular Cloning: A Laboratory Manual
    • Maniatis, T.1    Fritsch, E.F.2    Sambrook, J.3
  • 28
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    • + RNA for 2 hours at 60°C. The extension reaction was performed at 41°C for 1 hour with avian myeloblastosis virus (AMV) reverse transcriptase (Life Sciences, Inc.) and terminated by addition of NaOH to 0.4M and EDTA to 20 mM, with subsequent neutralization with ammonium acetate (pH 4.6). The products were treated with phenol, precipitated with ethanol, redissolved in buffer with formamide, and analyzed on a polyacrylamide sequencing gel
    • (1987) Methods Enzymol. , vol.152 , pp. 611
    • Calzone, F.J.1    Britten, R.J.2    Davidson, E.H.3
  • 30
    • 0022372670 scopus 로고
    • 12-3″. This linker, together with another primer (internal to the extension primer) with an Eco RI restriction site at its 5″ end, was then used for PCR. After digestion with Eco RI and Bgl II, products were purified and cloned in Bluescript KS (Stratagene) by standard procedures. All the recovered clones contained inserts of more than 350 nt. The 3″ end clones were generated with the use of similar procedures. PCR amplification was carried out with the linker described above and an oligonucleotide with the sequence 5″-ATGAAGTCCAAGGAnTAG-3″, which is ~70 nt upstream of the Hind III site at position 5027 (Fig. 2). The products were digested with Hind III and Xho I and cloned in the Bluescript vector. Candidate clones were identified by hybridization with the 3″ end of cDNA T16-4.5. All PCR's were performed for 30 cycles as described by the enzyme supplier
    • (1985) Science , vol.230 , pp. 1350
    • Saiki, R.K.1
  • 32
    • 0022552131 scopus 로고
    • (1986) Cell , vol.44 , pp. 283
  • 39
    • 0022166121 scopus 로고
    • Several large families of integral membrane proteins are known, including: (i) A number of ligand-gated ion channels of which the nicotinic acetylcholine receptor is the prototype
    • (1985) Annu. Rev. Cell Biol. , vol.1 , pp. 317
    • Stroud, R.M.1    Finer-Moore, J.2
  • 60
    • 0019423856 scopus 로고
    • + channel and CFTR. Similarities between sequences at the same relative locations with respect to the COOH-termini of the raf kinase and CFTR suggest that they may also share at least a small facet of their structures and functions
    • (1981) Nature , vol.290 , pp. 457
    • Anderson, S.1


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