Cloning and sequence of full‐length cDNAs encoding the human neuronal nicotinic acetylcholine receptor (nAChR) subunits β3 and β4 and expression of seven nAChR subunits in the human neuroblastoma cell line SH‐SY5Y and/or IMR‐32

FEBS Letters

Luyten, Walter H.M.L¹  Groot Kormelink, Paul J² 

[1] Experimental Molecular Biology Department, Janssen Research Foundation, Turnhoutseweg 30, B-2340 Beerse, Belgium;Laboratory for Molecular Biology, Celgen, University of Leuven, Herestraat 49, B-3000 Leuven, Belgium

关键词: Neuroblastoma;    Nicotinic receptor;    Polymerase chain reaction;    Gene expression;    Human;    nAChR;    nicotinic acetylcholine receptor;    TD-PCR;    touch-down polymerase chain reaction;    RACE;    rapid amplification of cDNA ends;    Tm;    melting temperature;    nt;    nucleotide;    bp;    base pair;    kb;    kilobase;    dNTP;    deoxynucleotide triphosphate;    UTR;    untranslated region;    GSP;    gene-specific primer;    AP;    adapter primer;    MEM;    modified Eagle's medium;    DMEM;    Dulbecco MEM;    DMSO;    dimethylsulphoxide;    SDS;    sodium dodecyl sulphate;    SSC;    standard sodium chloride/sodium citrate buffer;    SSPE;    standard sodium chloride/sodium phosphate/ethylene diamine tetraacetate buffer;   

DOI  :  10.1016/S0014-5793(96)01383-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Using PCR-based techniques, we have isolated and sequenced the full-length cDNAs that encode the human neuronal nAChR subunits α3,4,5,7 and β2,3,4 in the neuroblastoma cell lines SH-SY5Y and/or IMR-32. The predicted nAChR β3- and β4-subunit proteins contain 458 and 498 amino acids, respectively. Except for the β2, all the other cloned cDNAs showed differences with the published sequences. Northern blots show expression of the nAChR subunits α3,7 and β2,4 in the human neuroblastoma cell line SH-SY5Y, with intensity of the hybridisation signal decreasing in the order α3>β4>β2>α7.

【 授权许可】

Unknown