Perreau J, Lilienbaum A, Vasseur M, Paulin D
Université Paris 7, France.
Gene. 1988;62(1):7-16. doi: 10.1016/0378-1119(88)90575-6.
We have previously reported the identification and isolation by mRNA selection/translation of a recombinant clone containing 80% of the human vimentin gene sequence [Lilienbaum et al., EMBO J. 5 (1986) 2809-2814]. We present here the nucleotide sequence of this genomic clone including the 3' untranslated region. To complete the coding sequence, we have isolated cDNA recombinant clones (1.1 kb) of vimentin from human libraries constructed in lambda gt11. Comparison of the coding sequence between human and hamster shows 90% homology at the nucleotide level and four differences out of 353 amino acid residues, as deduced from the nucleotide sequences. In addition to the extensive homology previously reported between the coding sequences of hamster and human vimentin genes [Ferrari et al., Mol. Cell. Biol. 6 (1986) 3614-3620], we observed that the positions of the noncoding regions are also conserved and that the 3' nontranslated region includes two canonic poly(A) signals. Hybridization of the clones to mRNA from different mammalian sources revealed a single species of 2 kb and confirmed that the length of the untranslated and coding sequences are conserved. Quantitative estimations of the mRNA levels in mammalian cells and tissues of various origins are consistent with transcriptional regulation.
我们之前报道过通过mRNA筛选/翻译鉴定并分离出一个重组克隆,该克隆包含80%的人波形蛋白基因序列[利连鲍姆等人,《欧洲分子生物学组织杂志》5 (1986) 2809 - 2814]。我们在此展示该基因组克隆的核苷酸序列,包括3'非翻译区。为了完成编码序列,我们从构建于λgt11的人源文库中分离出波形蛋白的cDNA重组克隆(1.1 kb)。人与仓鼠编码序列的比较显示,在核苷酸水平上有90%的同源性,从核苷酸序列推导,在353个氨基酸残基中有四个差异。除了之前报道的仓鼠和人波形蛋白基因编码序列之间的广泛同源性[法拉利等人,《分子与细胞生物学》6 (1986) 3614 - 3620],我们还观察到非编码区的位置也保守,并且3'非翻译区包含两个典型的聚腺苷酸化信号。这些克隆与来自不同哺乳动物来源的mRNA杂交显示出一种2 kb的单一物种,并证实了非翻译区和编码序列的长度是保守的。对各种来源的哺乳动物细胞和组织中mRNA水平的定量估计与转录调控一致。