Ceratto N, Dobkin C, Carter M, Jenkins E, Yao X L, Cassiman J J, Aly M S, Bosco P, Leube R, Langbein L, Feo S, Romano V
Laboratorio di Genetica Molecolare, Istituto per la Ricerca sul Ritardo Mentale e l'Involuzione Cerebrale (I.R.C.C.S.), Troina, Italy.
Cytogenet Cell Genet. 1997;77(3-4):169-74. doi: 10.1159/000134566.
A YAC clone (211F11) containing approximately 0.5 Mb of human DNA was isolated from a human genomic library by PCR-based screening with cytokeratin (KRT) 13-specific primers. The YAC clone was mapped by FISH to the long arm of chromosome 17 (17q12-->q21), a region to which several other type I KRT genes had been mapped previously. We now show by Southern blot hybridization and PFGE analyses that KRT13, 14, 15, and 16 are all contained within YAC clone 211F11. Long-range restriction mapping analysis of clone 211F11 and of two smaller YAC clones that were also isolated with KRT13-specific primers, suggests that KRT13, 14, 15, 16 and their linked type I genes KRT17 and 19, are contained in less than 150 kb of genomic DNA. According to our reconstruction it then appears that at least six type I KRT genes are arranged in a highly compact cluster. The three YACs reported in this study represent a new tool to dissect the molecular structure of the locus of the human type I KRT genes.
通过使用细胞角蛋白（KRT）13特异性引物进行基于PCR的筛选，从人类基因组文库中分离出一个包含约0.5 Mb人类DNA的酵母人工染色体（YAC）克隆（211F11）。通过荧光原位杂交（FISH）将该YAC克隆定位到17号染色体的长臂（17q12→q21），之前已有几个其他I型KRT基因被定位到该区域。我们现在通过Southern印迹杂交和脉冲场凝胶电泳（PFGE）分析表明，KRT13、14、15和16都包含在YAC克隆211F11中。对克隆211F11以及另外两个也用KRT13特异性引物分离出的较小YAC克隆进行的长距离限制性图谱分析表明，KRT13、14、15、16及其连锁的I型基因KRT17和19包含在不到150 kb的基因组DNA中。根据我们的重构，至少六个I型KRT基因似乎排列成一个高度紧凑的簇。本研究中报道的三个YAC代表了一种剖析人类I型KRT基因位点分子结构的新工具。
