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大鼠脑中一种新型脂肪酸结合蛋白编码cDNA的克隆与鉴定

Cloning and characterization of a cDNA encoding a novel fatty acid binding protein from rat brain.

作者信息

Bennett E, Stenvers K L, Lund P K, Popko B

机构信息

Brain and Development Research Center, University of North Carolina at Chapel Hill 27599-7250.

出版信息

J Neurochem. 1994 Nov;63(5):1616-24. doi: 10.1046/j.1471-4159.1994.63051616.x.

Abstract

We have adopted a polymerase chain reaction approach to identify and clone a cDNA that contains the complete coding sequence of a novel fatty acid binding protein (FABP) from a rat brain lambda gt10 library. Sequencing of the brain FABP (B-FABP) cDNA revealed an open reading frame coding for a protein with 132 amino acids and a predicted size of approximately 15,000 Da. This putative protein shares extensive sequence homology with other members of the FABP family. Northern blot analysis using the B-FABP cDNA as a probe established the presence of an abundant mRNA approximately 0.8 kb long in rat brain and in the MOCH-1 oligodendrocyte cell line. This transcript was also present in rat liver but not in other tissues examined. A developmental profile of this mRNA in rat brain demonstrated detectable expression in 15-day-old embryos with levels peaking in 1-day postnatal neonates and declining thereafter, reaching a low steady-state level at 3 weeks of age. In situ hybridization histochemistry revealed B-FABP mRNA in various brain regions, with the highest levels in fiber tracts. The B-FABP message was also detected at a lower level in several gray matter regions. The cloning approach used in this study would likely be useful in the identification and isolation of FABP-encoding genes from other tissues and species.

摘要

我们采用聚合酶链反应方法,从大鼠脑λgt10文库中鉴定并克隆了一个包含新型脂肪酸结合蛋白(FABP)完整编码序列的cDNA。脑FABP(B-FABP)cDNA测序显示一个开放阅读框,编码一个含132个氨基酸、预测大小约为15,000 Da的蛋白质。这个假定的蛋白质与FABP家族的其他成员具有广泛的序列同源性。以B-FABP cDNA为探针进行Northern印迹分析,证实大鼠脑和MOCH-1少突胶质细胞系中存在一种约0.8 kb长的丰富mRNA。该转录本也存在于大鼠肝脏中,但在所检测的其他组织中不存在。大鼠脑中这种mRNA的发育表达谱显示,在15日龄胚胎中可检测到表达,出生后1天的新生鼠中表达水平达到峰值,此后下降,在3周龄时达到低稳态水平。原位杂交组织化学显示,B-FABP mRNA存在于脑的各个区域,在纤维束中水平最高。在几个灰质区域也检测到较低水平的B-FABP信息。本研究中使用的克隆方法可能有助于从其他组织和物种中鉴定和分离编码FABP的基因。

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