富集斑马鱼(Danio rerio) cDNA文库中N-ras原癌基因的克隆、测序和胚胎表达

R Cheng, S Bradford, D Barnes, D Williams, J Hendricks, G Bailey
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引用次数: 0

摘要

构建丰富的斑马鱼(Danio rerio)互补DNA文库,筛选ras相关基因,并从3 × 10(4)个斑块的一个板中分离出阳性克隆。该克隆为Zras-B1,内含2592个碱基对(bp)和一个开放阅读框,编码ras p21蛋白188个氨基酸。推导出的氨基端86个氨基酸残基和羧基端CAAX结合基序与哺乳动物ras相同。zras - b1编码的全长蛋白与人类N-ras的同源性最密切(同源性为91%),与Ha-ras的同源性次之(同源性为84%)和Ki-ras的同源性为85%)。初步筛选数据还表明斑马鱼中存在其他ras基因,其中至少有一种基因在成年斑马鱼中也有转录。一个与zras - b1相关的3.1 kb转录本在受精卵到原肠胚形成的胚胎中大量存在,可能来自母体。
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Cloning, sequencing, and embryonic expression of an N-ras proto-oncogene isolated from an enriched zebrafish (Danio rerio) cDNA library.

An enriched zebrafish (Danio rerio) complementary DNA library was constructed for screening of ras-related genes, and a positive clone was isolated from one plate of 3 x 10(4) plaques. This clone, Zras-B1, carried an insert of 2592 base pairs (bp) with an open reading frame encoding a ras p21 protein of 188 amino acids. The deduced amino-terminal 86 amino acid residues and the carboxy-terminal CAAX binding motif are identical to mammalian ras. The full-length Zras-B1-encoded protein is most closely related to human N-ras (91% identity), with lesser homology to Ha-ras (84%) and Ki-ras (85%). Preliminary screening data also indicate other ras genes in zebrafish, at least one of which is also transcribed in adults. A Zras-B1-related 3.1-kb transcript was found to be abundant in embryos from zygote through gastrulation, and may be maternally derived.

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