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小鼠鸟氨酸脱羧酶mRNA的长前导序列,以前被怀疑是克隆假象,可能是与MuLV样逆转录病毒重组的产物。

The long leader sequence of the mouse ornithine decarboxylase mRNA, previously suspected to be a cloning artifact, is probably a product of recombination with MuLV-like retrovirus.

作者信息

Widegren B, Blomberg J, Arnason U

机构信息

Department of Molecular Genetics, Wallenberg Laboratory, Lund, Sweden.

出版信息

Gene. 1991 Dec 30;109(2):303-5. doi: 10.1016/0378-1119(91)90626-m.

Abstract

Sequence analysis of the first 549 nucleotides (nt) of the non-translated 5' end of the cloned mouse ornithine decarboxylase (ODC; L-ornithine carboxy-lyase, EC 4.1.1.17)-encoding sequence shows that this sequence is closely related to nt 1946-1395 of Moloney murine leukemia virus (MuLV). The viral sequence, however, is oriented anti-sense relative to the ODC sequence. This orientation makes it unlikely to be a cloning artifact mediated by reverse transcriptase, but rather a recombination between genomic DNA and a MuLV-like provirus. In the cell line, from which the cDNA clone originated, Katz and Kahana [EMBO J. 8 (1989) 1163-1167] have shown that an intragenic deletion and amplification of the ODC gene had taken place. We believe that an additional recombination also has occurred in this cell line. The cDNA clone studied was obtained after selecting for high ODC expression. It is conceivable that the retroviral sequence contains an intragenic enhancer which is also functional in the anti-sense orientation. The inserted sequence contains two repeats which share homology with known enhancer elements. The reported recombination event shows that caution is needed when selective pressure is applied for the isolation and characterization of genes.

摘要

对克隆的小鼠鸟氨酸脱羧酶(ODC;L-鸟氨酸羧基裂解酶,EC 4.1.1.17)编码序列的非翻译5'端的前549个核苷酸(nt)进行序列分析表明,该序列与莫洛尼鼠白血病病毒(MuLV)的nt 1946 - 1395密切相关。然而,病毒序列相对于ODC序列呈反义方向。这种方向使得它不太可能是由逆转录酶介导的克隆假象,而更可能是基因组DNA与类似MuLV的前病毒之间的重组。在该cDNA克隆所源自的细胞系中,Katz和Kahana [《欧洲分子生物学组织杂志》8 (1989) 1163 - 1167] 已经表明ODC基因发生了基因内缺失和扩增。我们认为在这个细胞系中还发生了另外一次重组。所研究的cDNA克隆是在选择高ODC表达后获得的。可以想象,逆转录病毒序列包含一个基因内增强子,其在反义方向上也具有功能。插入序列包含两个与已知增强子元件具有同源性的重复序列。所报道的重组事件表明,在对基因进行分离和表征施加选择压力时需要谨慎。

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