Cloning and characterization of a mouse type I hair keratin cDNA.
Cloning and characterization of a mouse type I hair keratin cDNA.
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小鼠 I 型毛发角蛋白 cDNA 的克隆和表征。
DOI：
10.1111/1523-1747.ep12476901
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发表时间：
1988
期刊：
影响因子：
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通讯作者：
DiDona,GJ
中科院分区：
文献类型：
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作者：
Bertolino,AP;Checkla,DM;Notterman,R;Sklaver,I;Schiff,TA;Freedberg,IM;DiDona,GJ
以含Poly(A)的C57BL/6J小鼠毛根富集型mRNA为模板，构建了C57BL/6J小鼠毛根富集型mRNA文库。用从绵羊羊毛I型角蛋白基因克隆(SWK2)制备的放射性标记核酸探针对文库进行筛选。克隆MHKA-1经阳性杂交-选择翻译实验证明含有一条小鼠毛发、I型角蛋白基因插入片段，并推导出相应的氨基酸序列。该插入片段的大小为1585bp(不包括均聚体尾巴)，Northern印迹分析表明，该插入片段对应于一个约1.6kb的mRNA。克隆的cDNA序列包括416个氨基酸(包括蛋氨酸起始区)的完整编码区。将推导的氨基酸序列与绵羊羊毛I型角蛋白(8c1)的氨基酸序列进行比较，发现总的同源性为87%。相比之下，MHKA-1蛋白与非毛发I型角蛋白的相似性明显降低。在小鼠蛋白质的羧基末端的中间附近发现了另外3种不存在于羊毛蛋白中的氨基酸(相邻的脯氨酸残基)。MHKA-1是第一个被鉴定和鉴定的小鼠毛囊cDNA克隆，将使我们能够研究毛囊调节机制和哺乳动物毛囊中蛋白质之间的相互关系。
A cDNA library was prepared from poly(A)-containing C57BL/6J mouse hair-root-enriched mRNA. The library was screened using a radiolabeled nucleic acid probe prepared from a sheep wool, Type I keratin cDNA clone (SWK2). Clone MHKA-1 was shown to contain a mouse hair, Type I keratin cDNA insert by positive hybridization-selection translation assay, and by the corresponding deduced amino acid sequence. The size of the cDNA insert is 1585 bp (excluding homopolymer tails) and on the basis of Northern blot analysis it corresponds to a mRNA of approximately 1.6 Kb. The cloned cDNA sequence includes the entire coding region for a protein of 416 amino acids (including the initiation methionine). Comparison of the deduced amino acid sequence with that of sheep wool, Type I keratin (8c1) reveals an overall corresponding sequence identity of 87%. In contrast, the MHKA-1 protein is significantly less similar to non-hair Type I keratins. An additional 3 amino acids (adjacent proline residues) not present in the wool protein have been identified near the middle of the carboxy-terminal end of the mouse protein. MHKA-1 is the first of a series of mouse hair follicle cDNA clones to be identified and characterized that will enable us to study follicular regulatory mechanisms and the interrelationships among the proteins in the mammalian hair follicle.