Shimasaki S, Emoto N, Koba A, Mercado M, Shibata F, Cooksey K, Baird A, Ling N
Laboratories for Neuroendocrinology, Salk Institute, La Jolla, CA 92037.
Biochem Biophys Res Commun. 1988 Nov 30;157(1):256-63. doi: 10.1016/s0006-291x(88)80041-x.
Three cDNA clones encoding rat basic fibroblast growth factor (FGF) were isolated from 10(6) independent clones prepared from a pregnant mare serum gonadotropin (PMSG)-stimulated rat ovarian cDNA library. One of the cDNA clones contained the entire coding sequence for basic FGF. The other two possessed the sequence coding the carboxy terminal 61 amino acids of rat basic FGF, the putative upstream intron sequence, and a 3'-noncoding region. The cDNAs encoding rat basic FGF predict a molecule consisting of 154 amino acid residues, which is one amino acid shorter than the human and bovine basic FGF. Otherwise, there are only 5 conservative amino acid substitutions between the rat and the human/bovine sequences. Poly A+ RNA from brain cortex and hypothalamus show a single 6.0 kb band that hybridizes to the cloned cDNA probe by Northern analyses. The observation that basic FGF mRNA is below the limits of detection in adrenal, spleen, heart, lung, kidney, liver, stomach, small intestine, large intestine, testis, and ovary support the notion that the that the high levels of the protein found in these tissues is due to storage of the mitogen in the extracellular matrix and not continuous gene expression. The significance of the abundance of mRNA in tissues which are not undergoing either active angiogenesis or cell proliferation (hypothalamus and brain cortex) is unclear but emphasizes the potential neuronotrophic function of basic FGF.
从由孕马血清促性腺激素(PMSG)刺激的大鼠卵巢cDNA文库制备的10^6个独立克隆中,分离出了三个编码大鼠碱性成纤维细胞生长因子(FGF)的cDNA克隆。其中一个cDNA克隆包含碱性FGF的完整编码序列。另外两个拥有编码大鼠碱性FGF羧基末端61个氨基酸的序列、推定的上游内含子序列以及一个3'非编码区。编码大鼠碱性FGF的cDNAs预测一个由154个氨基酸残基组成的分子,该分子比人和牛的碱性FGF短一个氨基酸。除此之外,大鼠与人/牛序列之间仅有5个保守氨基酸替换。通过Northern分析,来自大脑皮层和下丘脑的聚腺苷酸加尾RNA(Poly A+ RNA)显示出一条与克隆的cDNA探针杂交的单一6.0 kb条带。在肾上腺、脾脏、心脏、肺、肾脏、肝脏、胃、小肠、大肠、睾丸和卵巢中未检测到碱性FGF mRNA,这一观察结果支持了这样一种观点,即这些组织中发现的高水平蛋白质是由于有丝分裂原储存在细胞外基质中,而不是由于持续的基因表达。在既没有活跃血管生成也没有细胞增殖的组织(下丘脑和大脑皮层)中mRNA丰度的意义尚不清楚,但强调了碱性FGF潜在的神经营养功能。