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大鼠中一种叶酸结合蛋白基因的编码序列、基因组结构及表达

Coding sequence, genomic organization and expression of a folate binding protein gene in the rat.

作者信息

Sadasivan E, Meng Y, Rothenberg S P

机构信息

Division of Hematology/Oncology State University of New York-Downstate Medical Center, 450 Clarkson Avenue - Box 20, Brooklyn, NY 11203, USA.

出版信息

Gene. 2000 Aug 22;254(1-2):219-28. doi: 10.1016/s0378-1119(00)00262-6.

Abstract

The complementary DNA (cDNA) and the gene encoding the folate binding protein alpha isoform (FBPalpha) have been reported for the human and mouse protein. However, there is no information about this gene in the rat, an animal that could be a model to study expression of this protein in vivo when folate metabolism is modified. Accordingly, the cloning and characterization of this gene in the rat have been the subject of this research. The gene has seven exons and six introns and is approximately 10kb in size. The organization and nucleotide sequence of the coding exons are similar to those of the corresponding human and mouse genes, which are the only other mammalian FBP genes cloned. However, the amino acid sequence of the rat FBPalpha is less homologous, having 48% identity with the published sequences for all the mammalian FBP isoforms. A finding not previously reported is the expression of two FBPalpha transcripts in the kidney that differ in the length of the 5' untranslated sequences, as determined by rapid amplification of cDNA end-polymerase chain reaction amplification (RACE-PCR). The brain expresses a single transcript intermediate in size between the two transcripts expressed in the kidney. The kidney transcripts are encoded by the same gene and appear to be regulated either from two independent promoters or from a single promoter in association with alternative RNA splicing.

摘要

人类和小鼠叶酸结合蛋白α亚型(FBPα)的互补DNA(cDNA)及编码该蛋白的基因已见报道。然而,对于大鼠这一动物模型,当叶酸代谢发生改变时,其体内该蛋白表达情况的相关研究中却并无此基因的信息。因此,本研究的主题是大鼠中该基因的克隆与特性分析。该基因有7个外显子和6个内含子,大小约为10kb。编码外显子的结构和核苷酸序列与已克隆的仅有的另外两种哺乳动物FBP基因,即相应的人类和小鼠基因相似。然而,大鼠FBPα的氨基酸序列同源性较低,与所有已发表的哺乳动物FBP亚型序列的一致性仅为48%。一项此前未报道的发现是,通过cDNA末端快速扩增-聚合酶链反应扩增(RACE-PCR)测定,在肾脏中表达的两种FBPα转录本在5'非翻译序列长度上存在差异。大脑表达的单一转录本大小介于肾脏中表达的两种转录本之间。肾脏转录本由同一基因编码,似乎是由两个独立启动子或与可变RNA剪接相关的单一启动子调控。

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