Identification of five new genes on the Y chromosome of Drosophila melanogaster
Identification of five new genes on the Y chromosome of Drosophila melanogaster
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DOI：
10.1073/pnas.231484998
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发表时间：
2001-11-06
影响因子：
11.1
通讯作者：
Clark, AG
中科院分区：
文献类型：
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作者：
Carvalho, AB;Dobo, BA;Clark, AG
果蝇Y染色体的异染色质状态使得Y连锁基因的克隆和鉴定成为一个具有挑战性的过程。在这里，我们报告的程序，以确定Y-连锁的基因片段从整个基因组测序工作的未映射的残基，以前确定的Y-连锁的基因出现在测序的支架作为单独的外显子，显然是因为许多内含子已成为异染色质，增长到巨大的尺寸，变得几乎不可克隆。使用所有已知的蛋白质作为查询序列，对未映射片段的可爆破数据库进行测试，TBLAYBLOG搜索产生了与这种情况一致的许多匹配。逆转录-PCR和遗传学方法用于确认那些表达的Y连锁基因。本文报道的5个基因包括3个蛋白磷酸酶（Pp 1-Y1、Pp 1-Y2和PPr-Y）、1个闭合蛋白相关基因（ORY）和1个卷曲螺旋基因（CCY）。这使得果蝇Y染色体上总共有9个蛋白质编码基因。ORY和CCY可能分别对应于生育因子ks-1和ks-2，而三种蛋白磷酸酶代表新基因。果蝇Y染色体上的基因之间仍然存在着与雄性功能的强烈功能一致性。
The heterochromatic state of the Drosophila Y chromosome has made the cloning and identification of Y-linked genes a challenging process. Here, we report application of a procedure to identify Y-linked gene fragments from the unmapped residue of the whole genome sequencing effort, Previously identified Y-linked genes appear in sequenced scaffolds as individual exons apparently because many introns have become heterochromatic, growing to enormous size and becoming virtually unclonable. A TBLASTN search using all known proteins as query sequences, tested against a blastable database of the unmapped fragments, produced a number of matches consistent with this scenario. Reverse transcription-PCR and genetic methods were used to confirm those that are expressed, Y-linked genes. The five genes reported here include three protein phosphatases (Pp1-Y1, Pp1-Y2, and PPr-Y), an occludin-related gene (ORY), and a coiled-coils gene (CCY). This brings the total to nine protein-coding genes identified on the Drosophila Y chromosome. ORY and CCY may correspond, respectively, to the fertility factors ks-1 and ks-2, whereas the three protein phosphatases represent novel genes. There remains a strong functional coherence to male function among the genes on the Drosophila Y chromosome.