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通过外显子捕获从牛黏粒中分离编码序列。

Isolation of coding sequences from bovine cosmids by means of exon trapping.

作者信息

Comincini S, Drisaldi B, Ferretti L

机构信息

Istituto per la Difesa e la Valorizzazione del Germoplasma Animale (I.D.V.G.A.) - C.N.R., via Celoria 10, 20133 Milano, Italy.

出版信息

Mamm Genome. 1997 Jul;8(7):486-90. doi: 10.1007/s003359900481.

Abstract

Exon trapping was employed to identify coding sequences from a collection of 46 bovine cosmids, previously characterized for the presence of microsatellite markers and physically mapped to chromosomes by FISH. The sequence analysis of 104 clones revealed 18 putative exons, 10 of which showed near identity to known sequences. Among these were the human (cytosine-5)-methyltransferase (DNMT), ATP-citrate lyase (ACLY), the mouse Lbcl1 oncogene, the bovine mitochondrial aconitase (ACO2) and beta-arrestin 1 (ARR1). The chromosomal localization of the cloned exons was inferred from the localization of the parent cosmids. DNMT and ACLY were not previously known in cattle, but the physical localization of the cloned bovine exons is in agreement with the published comparative human and bovine maps. The trapping of exons for bovine ACO2 and ARR1 confirms the available mapping information based on synteny and provides a physical assignment for the genes.

摘要

外显子捕获技术被用于从46个牛黏粒文库中鉴定编码序列,这些黏粒先前已通过微卫星标记进行了特征鉴定,并通过荧光原位杂交(FISH)技术进行了染色体物理定位。对104个克隆的序列分析揭示了18个推定的外显子,其中10个与已知序列高度相似。其中包括人类(胞嘧啶-5)-甲基转移酶(DNMT)、ATP-柠檬酸裂解酶(ACLY)、小鼠Lbcl1癌基因、牛线粒体乌头酸酶(ACO2)和β-抑制蛋白1(ARR1)。克隆外显子的染色体定位是根据亲本黏粒的定位推断出来的。DNMT和ACLY在牛中以前并不为人所知,但克隆的牛外显子的物理定位与已发表的人类和牛的比较图谱一致。牛ACO2和ARR1外显子的捕获证实了基于同线性的现有定位信息,并为这些基因提供了物理定位。

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