Mariman E C, Broers C A, Claesen C A, Tesser G I, Wieringa B
Department of Human Genetics, University Nijmegen, Radboud Hospital, The Netherlands.
Genomics. 1987 Oct;1(2):126-37. doi: 10.1016/0888-7543(87)90004-8.
Various cDNAs for creatine kinase type B (CK-B) were isolated from human cDNA libraries using a 26-oligonucleotide guess-mer probe. One of the cDNAs appeared to be almost full-length and contained an open reading frame coding for the 381 amino acid residues of the human CK-B polypeptide. The nucleotide sequences of the translated region as well as the primary protein structure show a high degree of homology with known CK-B and CK-M sequences of other vertebrates. The level of CK-B RNA as a measure of CK-B gene activity was determined in various human tissues and cultured cells. Our results confirm that CK-B is expressed in a tissue-specific manner and give support to the previously proposed relation between CK-B gene activity and cell proliferation. Screening of genomic DNA with various cDNA regions as probes revealed that there is only one CK-B gene per haploid genome. Gene cloning and sequencing indicated that CK-B is coded for by a relatively small gene of 3.2 kb in size, which is partially overlapped by an HTF island (A. P. Bird (1986) Nature (London) 321, 557-558) with an extremely high G + C content at its 5' end.
使用一个26个寡核苷酸的推测性探针,从人cDNA文库中分离出了各种肌酸激酶B型(CK-B)的cDNA。其中一个cDNA似乎几乎是全长的,包含一个编码人CK-B多肽381个氨基酸残基的开放阅读框。翻译区域的核苷酸序列以及蛋白质一级结构与其他脊椎动物已知的CK-B和CK-M序列显示出高度同源性。在各种人体组织和培养细胞中测定了作为CK-B基因活性指标的CK-B RNA水平。我们的结果证实CK-B以组织特异性方式表达,并支持了先前提出的CK-B基因活性与细胞增殖之间的关系。用各种cDNA区域作为探针筛选基因组DNA表明,每个单倍体基因组只有一个CK-B基因。基因克隆和测序表明,CK-B由一个相对较小的3.2 kb基因编码,该基因在其5'端部分与一个G + C含量极高的HTF岛重叠(A. P. Bird(1986年),《自然》(伦敦)321,557 - 558)。