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胶质细胞成熟因子β mRNA和蛋白在大鼠器官及细胞中的表达

Expression of glia maturation factor beta mRNA and protein in rat organs and cells.

作者信息

Zaheer A, Fink B D, Lim R

机构信息

Department of Neurology, Division of Neurochemistry and Neurobiology, University of Iowa College of Medicine, Iowa City 52242.

出版信息

J Neurochem. 1993 Mar;60(3):914-20. doi: 10.1111/j.1471-4159.1993.tb03237.x.

Abstract

Rat glia maturation factor beta (GMF-beta) cDNA was obtained by reverse transcription of rat brain mRNA followed by polymerase chain reaction amplification, using primers from the human sequence. The deduced amino acid sequence of rat GMF-beta differed from the human counterpart in only three places: His27 in place of Asn, Val51 in place of Ile, and Leu93 in place of Val. The high degree of evolutionary conservation suggests that GMF-beta plays an essential role in animal cell physiology. The expression of GMF-beta mRNA in the rat was studied by the northern blot technique, using a rat cRNA probe corresponding to the entire coding region. GMF-beta mRNA was predominantly expressed in the brain and spinal cord, although trace levels were found in other organs, including testis and ovary. In the brain GMF-beta mRNA was detectable at as early as embryonic day 10, and persisted through as late as postnatal month 14, with minor variations in between. On the other hand, GMF-beta protein exhibited more obvious developmental changes, with its level increasing slowly prenatally and plateauing at 1 week after birth. GMF-beta mRNA and protein were also observed in several cultured cells. Some cells of neural origin contained higher levels of GMF-beta protein compared with cells derived from other sources. Through demonstration of mRNA and confirmation by immunoblotting, we conclude that GMF-beta is synthesized by rat organs and that GMF-beta is predominantly a brain protein.

摘要

大鼠胶质细胞成熟因子β(GMF-β)cDNA是通过逆转录大鼠脑mRNA,随后使用来自人类序列的引物进行聚合酶链反应扩增而获得的。大鼠GMF-β推导的氨基酸序列与人类对应序列仅在三个位置不同:His27取代Asn、Val51取代Ile以及Leu93取代Val。高度的进化保守性表明GMF-β在动物细胞生理学中发挥着重要作用。使用对应于整个编码区的大鼠cRNA探针,通过Northern印迹技术研究了大鼠中GMF-βmRNA的表达。GMF-βmRNA主要在脑和脊髓中表达,尽管在包括睾丸和卵巢在内的其他器官中发现了微量水平。在脑中,GMF-βmRNA早在胚胎第10天就可检测到,并一直持续到出生后第14个月,期间有微小变化。另一方面,GMF-β蛋白表现出更明显的发育变化,其水平在产前缓慢增加,并在出生后1周达到稳定状态。在几种培养细胞中也观察到了GMF-βmRNA和蛋白。与其他来源的细胞相比,一些神经源性细胞含有更高水平的GMF-β蛋白。通过mRNA的证明和免疫印迹的确认,我们得出结论,GMF-β由大鼠器官合成,并且GMF-β主要是一种脑蛋白。

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