Kaye F J, McBride O W, Battey J F, Gazdar A F, Sausville E A
J Clin Invest. 1987 May;79(5):1412-20. doi: 10.1172/JCI112969.
Using a small cell lung cancer (SCLC) cDNA library, we obtained clones for the creatine kinase-B (CK-B) gene and determined the nucleotide sequence for the protein coding and 3' untranslated region (3' UT). The human translated protein spans 381 residues and the amino acid homology with rabbit CK-B is greater than 98%. We have demonstrated that a nucleic acid probe encompassing the protein coding region will also hybridize to CK-M sequences while a probe derived from the 3' UT region is CK-B specific. When a B-isoenzyme specific sequence is hybridized to Eco RI cut genomic DNA, two independent restriction fragment polymorphisms are detected. We have subsequently localized these two CK-B homologous sequences to chromosomes 14q32 and 16. Finally, we show that increased levels of CK-B seen in SCLC are not accompanied by gene amplification or rearrangement, but reflect a greatly enhanced level of CK-B specific mRNA that is not seen in non-SCLC lines thus far examined.
利用一个小细胞肺癌(SCLC)cDNA文库,我们获得了肌酸激酶-B(CK-B)基因的克隆,并确定了蛋白质编码区和3'非翻译区(3'UT)的核苷酸序列。人翻译后的蛋白质跨度为381个残基,与兔CK-B的氨基酸同源性大于98%。我们已经证明,包含蛋白质编码区的核酸探针也会与CK-M序列杂交,而来自3'UT区的探针是CK-B特异性的。当一个B同工酶特异性序列与Eco RI切割的基因组DNA杂交时,可检测到两个独立的限制性片段多态性。我们随后将这两个CK-B同源序列定位到染色体14q32和16上。最后,我们表明,在SCLC中看到的CK-B水平升高并非伴随着基因扩增或重排,而是反映了CK-B特异性mRNA水平的大幅提高,这在迄今为止检测的非SCLC细胞系中未见。