Rogers M A, Langbein L, Winter H, Ehmann C, Praetzel S, Korn B, Schweizer J
Divisions of Biochemistry of Tissue-specific Regulation, Cell Biology, and Resource Center for Human Genome Research, German Cancer Research Center, 69120 Heidelberg, Germany.
J Biol Chem. 2001 Jun 1;276(22):19440-51. doi: 10.1074/jbc.M100657200. Epub 2001 Feb 27.
Low stringency screening of a human P1 artificial chromosome library using a human hair keratin-associated protein (hKAP1.1A) gene probe resulted in the isolation of six P1 artificial chromosome clones. End sequencing and EMBO/GenBank(TM) data base analysis showed these clones to be contained in four previously sequenced human bacterial artificial chromosome clones present on chromosome 17q12-21 and arrayed into two large contigs of 290 and 225 kilobase pairs (kb) in size. A fifth, partially sequenced human bacterial artificial chromosome clone data base sequence overlapped and closed both of these contigs. One end of this 600-kb cluster harbored six gene loci for previously described human type I hair keratin genes. The other end of this cluster contained the human type I cytokeratin K20 and K12 gene loci. The center of the cluster, starting 35 kb downstream of the hHa3-I hair keratin gene, contained 37 genes for high/ultrahigh sulfur hair keratin-associated proteins (KAPs), which could be divided into a total of 7 KAP multigene families based on amino acid homology comparisons with previously identified sheep, mouse, and rabbit KAPs. To date, 26 human KAP cDNA clones have been isolated through screening of an arrayed human scalp cDNA library by means of specific 3'-noncoding region polymerase chain reaction probes derived from the identified KAP gene sequences. This screening also yielded four additional cDNA sequences whose genes were not present on this gene cluster but belonged to specific KAP gene families present on this contig. Hair follicle in situ hybridization data for single members of five different KAP multigene families all showed localization of the respective mRNAs to the upper cortex of the hair shaft.
使用人发角蛋白相关蛋白（hKAP1.1A）基因探针，对人类P1人工染色体文库进行低严谨度筛选，分离出6个P1人工染色体克隆。末端测序及EMBO/GenBank数据库分析表明，这些克隆包含在4个先前已测序的人类细菌人工染色体克隆中，这些克隆位于17号染色体q12 - 21区域，并排列成两个大小分别为290和225千碱基对（kb）的大重叠群。第5个部分测序的人类细菌人工染色体克隆数据库序列与这两个重叠群都有重叠并使其封闭。这个600 kb簇的一端包含6个先前描述的人类I型毛发角蛋白基因的基因座。该簇的另一端包含人类I型细胞角蛋白K20和K12基因座。该簇的中心，从hHa3 - I毛发角蛋白基因下游35 kb处开始，包含37个高/超高硫毛发角蛋白相关蛋白（KAPs）的基因，根据与先前鉴定的绵羊、小鼠和兔KAPs的氨基酸同源性比较，这些基因可总共分为7个KAP多基因家族。迄今为止，通过使用从已鉴定的KAP基因序列衍生的特异性3'非编码区聚合酶链反应探针筛选排列好的人类头皮cDNA文库，已分离出26个人类KAP cDNA克隆。该筛选还产生了另外4个cDNA序列，其基因不在这个基因簇上，但属于这个重叠群上存在的特定KAP基因家族。五个不同KAP多基因家族的单个成员的毛囊原位杂交数据均显示各自的mRNA定位于毛干的上皮质。
