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采用逆转录-聚合酶链反应扩增法检测大鼠卵巢中酸性成纤维细胞生长因子mRNA。

Detection of acidic fibroblast growth factor mRNA in the rat ovary using reverse transcription-polymerase chain reaction amplification.

作者信息

Koos R D, Seidel R H

机构信息

Department of Physiology, University of Maryland School of Medicine, Baltimore 21201.

出版信息

Biochem Biophys Res Commun. 1989 Nov 30;165(1):82-8. doi: 10.1016/0006-291x(89)91036-x.

Abstract

We have examined the expression of acidic fibroblast growth factor (aFGF) in the rat ovary using reverse transcription-polymerase chain reaction (RT-PCR). RNA was extracted from hypothalami of adult rats and whole ovaries or isolated granulosa cells of gonadotropin-primed immature rats. The RNA was reverse transcribed and amplified by PCR using oligonucleotide primers specific for rat aFGF. RNA from hypothalamus or whole ovary yielded a dominant DNA band corresponding in size to the aFGF segment spanned by the two primers (301 base pairs, bp). Its identity with the aFGF sequence was confirmed by restriction enzyme analysis. The aFGF product was also obtained from two of four granulosa cell RNA preparations; when obtained, the intensity of the signal was less than that from whole ovary, indicating that the major sites of aFGF expression are outside the granulosa layer.

摘要

我们使用逆转录聚合酶链反应(RT-PCR)检测了大鼠卵巢中酸性成纤维细胞生长因子(aFGF)的表达。从成年大鼠的下丘脑以及促性腺激素预处理的未成熟大鼠的整个卵巢或分离的颗粒细胞中提取RNA。使用针对大鼠aFGF的寡核苷酸引物对RNA进行逆转录并通过PCR扩增。来自下丘脑或整个卵巢的RNA产生了一条占主导地位的DNA条带,其大小与两条引物跨越的aFGF片段相对应(301个碱基对,bp)。通过限制性酶切分析证实了其与aFGF序列的一致性。在四份颗粒细胞RNA制剂中的两份中也获得了aFGF产物;当获得该产物时,信号强度低于来自整个卵巢的信号强度,这表明aFGF表达的主要部位在颗粒层之外。

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