ISOLATION AND SEQUENCING OF A CDNA CODING FOR THE HUMAN DF3 BREAST CARCINOMA-ASSOCIATED ANTIGEN
ISOLATION AND SEQUENCING OF A CDNA CODING FOR THE HUMAN DF3 BREAST CARCINOMA-ASSOCIATED ANTIGEN
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DOI：
10.1073/pnas.85.7.2320
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发表时间：
1988-04-01
影响因子：
11.1
通讯作者：
KUFE, D
中科院分区：
文献类型：
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作者：
SIDDIQUI, J;ABE, M;KUFE, D
鼠单克隆抗体（mAb）DF 3与人乳腺癌中可检测到的高分子量糖蛋白反应。DF 3抗原表达与人乳腺肿瘤分化相关，并且在人乳中检测到交叉反应性物种表明该抗原可能用作分化的乳腺上皮的标志物。为了进一步表征DF 3抗原表达，我们通过用mAb DF 3筛选从λ gt 11文库中分离了cDNA克隆。结果表明，这个309个碱基对的cDNA，命名为pDF9.3，编码DF 3 spitope。用~（32）P标记的pDF9.3对6个人肿瘤细胞系的DNA进行Southern印迹分析，发现其限制性片段长度多态性，并在Pst Ⅰ中检测到等位基因大小的变异，而在HindIII中未检测到。此外，32 P标记的pDF9.3与来自这些细胞系中的每一个的总细胞RNA的杂交证明了一个或两个大小从4.1至7.1个碱基变化的转录本。通过pDF9.3检测到的不同大小的转录本的存在也被发现与DF 3糖蛋白的多态性表达相对应。pDF9.3的核苷酸序列分析揭示了一个高度保守的富含（G + C）的60碱基对串联重复序列。这些发现表明，多态性F3糖蛋白编码的等位基因的大小的变化可能代表不同数量的重复。
The murine monoclonal antibody (mAb) DF3 reacts with a high molecular weight glycoprotein detectable in human breast carcinomas. DF3 antigen expression correlates with human breast tumor differentiation, and the detection of a cross-reactive species in human milk has suggested that this antigen might be useful as a marker of differentiated mammary epithelium. To further characterize DF3 antigen expression, we have isolated a cDNA clone from a .lambda.gt11 library by screening with mAb DF3. The results demonstrate that this 309-base-pair cDNA, designated pDF9.3, codes for the DF3 spitope. Southern blot analyses of EcoRI-digested DNAs from six human tumor cell lines with 32P-labeled pDF9.3 have revealed a restriction fragment length polymorphism.Variations in size of the alleles detected by pDF9.3 were also identified in Pst I, but not in HindIII, DNA digests. Furthermore, hybridization of 32P-labeled pDF9.3 with total cellular RNA from each of these cell lines demonstrated either one or two transcripts that varied from 4.1 to 7.1 kilobases in size. The presence of differently size transcripts detected by pDF9.3 was also found to correspond with the polymorphic expression of DF3 glycoproteins. Nucleotide sequence analysis of pDF9.3 has revealed a highly conserved (G + C)-rich 60-base-pair tandem repeat. These findings suggest that the variation in size of alleles coding for the polymorphic F3 glycoprotein may represent different numbers of repeats.