Molecular analysis of the DNA segments cross-hybridizable to the tyrosinase gene in patients affected with oculocutaneous albinism.
Molecular analysis of the DNA segments cross-hybridizable to the tyrosinase gene in patients affected with oculocutaneous albinism.
复制标题
对眼皮肤白化病患者中可与酪氨酸酶基因交叉杂交的 DNA 片段进行分子分析。
DOI：
10.1620/tjem.159.333
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
S. Shibahara
中科院分区：
文献类型：
--
作者：
A. Takeda;J. Matsunaga;Y. Tomita;H. Tagami;S. Shibahara
人酪氨酸酶基因大于35 kb，并且被组织成四个内含子和五个外显子[Tomita等人（1989）Biochem.Biophys.Biophys.通信报告：164，990-996]。使用全长cDNA编码的人酪氨酸酶和其外显子特异性片段作为杂交探针，我们表明，酪氨酸酶基因的整体结构组织是不变的酪氨酸酶阴性眼皮肤白化病（OCA）的三名患者。此外，我们能够显示存在额外的DNA片段交叉杂交的外显子4或外显子5的酪氨酸酶基因的基因组中的三个OCA患者和一个健康个体。即，外显子4特异性探针在用EcoRI或HindIII消化的DNA中检测到两条带，外显子5特异性探针在Bgl II消化的DNA中检测到两条带，尽管事实上在每个外显子中不存在这些酶的识别位点。然后，我们分离了两个噬菌体克隆，它们含有不同的DNA片段，可与外显子5探针杂交。用Bgl II消化的每个克隆DNA的Southern印迹分析显示出与用Bgl II消化的基因组DNA中检测到的杂交模式不同的两种杂交模式，证实至少有两个DNA片段与人基因组中的外显子5序列杂交。
The human tyrosinase gene is greater than 35 kb and is organized in four introns and five exons [Tomita et al. (1989) Biochem. Biophys. Res. Commun., 164, 990-996]. Using the full-length cDNA encoding human tyrosinase and its exon-specific fragments as hybridization probes, we show that overall structural organization of the tyrosinase gene is unchanged in three patients affected with tyrosinase-negative oculocutaneous albinism (OCA). Moreover, we are able to show the presence of additional DNA segments cross-hybridizable to exon 4 or exon 5 of the tyrosinase gene in the genome of three OCA patients and a healthy individual. Namely, the exon 4 specific probe detected two bands in the DNA digested with EcoRI or HindIII and the exon 5-specific probe detected two bands in the Bgl II-digested DNA, in spite of the facts that no recognition sites for these enzymes are present in each exon. We have then isolated two phage clones harboring the distinct DNA segments, hybridizable to the exon 5 probe. Southern blotting analysis of each cloned DNA digested with Bgl II showed two different hybridization patterns as those detected in genomic DNA digested with Bgl II, confirming that there are at least two DNA segments hybridizing to the exon 5 sequence in human genome.
DOI：
10.1073/pnas.84.21.7473
发表时间：
1987-11-01
影响因子：
11.1
作者：
KWON, BS;HAQ, AK;HALABAN, R
通讯作者：
HALABAN, R