THE HUMAN GLUCOCEREBROSIDASE GENE AND PSEUDOGENE - STRUCTURE AND EVOLUTION
THE HUMAN GLUCOCEREBROSIDASE GENE AND PSEUDOGENE - STRUCTURE AND EVOLUTION
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DOI：
10.1016/0888-7543(89)90319-4
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发表时间：
1989-01-01
期刊：
影响因子：
4.4
通讯作者：
BEUTLER, E
中科院分区：
文献类型：
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作者：
HOROWITZ, M;WILDER, S;BEUTLER, E
我们报告了编码β-葡萄糖脑苷脂酶的整个人类基因的序列以及相关假基因的序列。该基因在整个序列中包含从碱基对355延伸至碱基对7232的11个外显子。基因启动子在葡萄糖脑苷脂酶 RNA 的主要 5'' 末端上游包含 TATA 和 CAT 样盒。两个 TATA 盒位于核苷酸 (-23)-(-27) 和 (-33)-(-39) 之间，两个可能的 CAT 盒位于与 mRNA 主要 5'' 末端相关的核苷酸 (-90)-(-94) 和 (-96)-(-99) 之间。通过将启动子区域与编码氯霉素乙酰转移酶（CAT）的细菌基因偶联并测定用该载体转染的细胞中该酶的表达来监测启动子区域的功能。葡萄糖脑苷脂酶启动子不仅指导细菌酶的合成，而且表现出与内源基因相同的组织特异性表达模式。明显紧密连锁的假基因与功能基因大约有 96% 同源性。然而，内含子 2、4、6 和 7 具有较大的“缺失”，分别由长度为 313、626、320 和 277 bp 的 Alu 序列组成。祖先基因完全有可能缺乏这些序列，并且它们已被插入到功能基因的内含子中。外显子 9 的一部分也有 55 bp 的缺失，两侧是短的反向重复序列。序列数据应有助于开发在分子水平上诊断戈谢病的方法。
We report the sequence of the entire human gene encoding .beta.-glucocerebrosidase and that of the associated pseudogene. The gene contains 11 exons extending from base pair 355 to base pair 7232 in the overall sequence. The gene promoter contains TATA-and CAT-like boxes upstream of the major 5'' end of the glucocerebrosidase RNA. The two TATA boxes lie between nucleotides (-23)-(-27) and (-33)-(-39) and the two possible CAT boxes reside between nucleotides (-90)-(-94) and (-96)-(-99) in relation to the major 5'' end of the mRNA. The functionality of the promoter region was monitored by coupling it to the bacterial gene coding for chloramphenicol acetyltransferase (CAT) and assaying the expression of the enzyme in cells transfected with this vector. The glucocerebrosidase promoter not only directs synthesis of the bacterial enzyme but also exhibits the same pattern of tissue-specific expression as that of the endogenous gene. An apparently tightly linked pseudogene is approximately 96% homologous to the functional gene. However, introns 2, 4, 6, and 7 have large "deletions" consisting of Alu sequences 313, 626, 320, and 277 bp in length, respectively. It is entirely possible that the ancestral gene lacks these sequences and that they have been inserted into the introns of the functioning gene. There is also a 55-bp deletion from a part of exon 9 flanked by a short inverted repeat. The sequence data should facilitate development of methods for diagnosis of Gaucher disease at the molecular level.