Van den Ouweland A M, Van Duijnhoven H L, Deichmann K A, Van Groningen J J, de Leij L, Van de Ven W J
Department of Biochemistry, University of Nijmegen, The Netherlands.
Nucleic Acids Res. 1989 May 25;17(10):3829-43. doi: 10.1093/nar/17.10.3829.
The monoclonal antibody MOC-32 detected a 40 kDa protein in Western blot analysis. Immunological screening of an expression library of human SCLC cells with MOC-32 led to the isolation of overlapping cDNA clones. One of these, cHD4, was 1.0 kbp long and of about the same size as its corresponding mRNA. Preceded by an in phase stop codon, an open reading frame of 885 bp was present in cHD4 and a translational product of only 33 kDa could be calculated. Biochemical and immunological analysis established the relationship between the 40 kDa antigen and the isolated coding sequences and resolved the apparent discrepancy between the calculated molecular weight and the observed electrophoretic mobility. Nucleotide sequence comparison of cHD4 to the EMBL database revealed that cHD4 was nearly identical to a sequence claimed to encode a laminin binding protein. Southern blot and nucleotide sequence analysis indicated the presence of multiple copies of the gene in the human genome. At least five of these appeared to represent processed pseudogenes.
单克隆抗体MOC - 32在蛋白质印迹分析中检测到一种40 kDa的蛋白质。用MOC - 32对人小细胞肺癌（SCLC）细胞的表达文库进行免疫筛选，导致分离出重叠的cDNA克隆。其中一个克隆cHD4长1.0 kbp，与其相应的mRNA大小相近。在cHD4中，一个885 bp的开放阅读框位于一个同相位终止密码子之前，并且可以计算出一个仅33 kDa的翻译产物。生化和免疫分析确定了40 kDa抗原与分离的编码序列之间的关系，并解决了计算分子量与观察到的电泳迁移率之间明显的差异。将cHD4的核苷酸序列与EMBL数据库进行比较，发现cHD4与一个声称编码层粘连蛋白结合蛋白的序列几乎相同。Southern印迹和核苷酸序列分析表明该基因在人类基因组中存在多个拷贝。其中至少有五个似乎代表加工过的假基因。
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