ISOLATION AND SEQUENCE OF A CDNA CLONE FOR HUMAN TYROSINASE THAT MAPS AT THE MOUSE C-ALBINO LOCUS
ISOLATION AND SEQUENCE OF A CDNA CLONE FOR HUMAN TYROSINASE THAT MAPS AT THE MOUSE C-ALBINO LOCUS
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DOI：
10.1073/pnas.84.21.7473
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发表时间：
1987-11-01
影响因子：
11.1
通讯作者：
HALABAN, R
中科院分区：
文献类型：
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作者：
KWON, BS;HAQ, AK;HALABAN, R
用抗仓鼠酪氨酸酶(单酚，L-多巴：氧氧化还原酶，EC 1.14.18.1)的抗体筛选λgt11人黑素细胞cDNA文库导致分离出16个克隆。 16 个克隆中有 13 个的 cDNA 插入片段相互交叉杂交，表明它们来自相关的 mRNA 物种。其中一个 cDNA 克隆 Pme134 检测到一种长度约为 2.4 kb 的 mRNA 种类，该 mRNA 优先在正常和恶性黑素细胞中表达，但不在其他细胞类型中表达。从核苷酸序列推导的氨基酸序列显示，推定的人酪氨酸酶由548个氨基酸组成，分子量为62,610。推导的蛋白质含有糖基化位点和富含组氨酸的位点，可用于铜结合。对携带致死性白化病缺失突变的新生小鼠的 DNA 进行 Southern 印迹分析，结果显示 Pme134 定位在 c-白化病基因座附近或该基因座上，即酪氨酸酶结构基因的位置。
Screening of a .lambda.gt11 human melanocyte cDNA library with antibodies against hamster tyrosinase (monophenol, L-dopa:oxygen oxidoreductase, EC 1.14.18.1) resulted in the isolation of 16 clones. The cDNA inserts from 13 of the 16 clones cross-hybridized with each other, indicating that they were from related mRNA species. One of the cDNA clones, Pme134, detected one mRNA species with an approximate length of 2.4 kilobases that was expressed preferentially in normal and malignant melanocytes but not in other cell types. The amino acid sequence deduced from the nucleotide sequence showed that the putative human tyrosinase is composed of 548 amino acids with a molecular weight of 62,610. The deduced protein contains glycosylation sites and histidine-rich sites that could be used for copper binding. Southern blot analysis of DNA derived from newborn mice carrying lethal albino deletion mutations revealed that Pme134 maps near or at the c-albino locus, the position of the structural gene for tyrosinase.