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增殖细胞核抗原的小鼠基因及假基因的分子克隆与结构分析

Molecular cloning and structural analysis of mouse gene and pseudogenes for proliferating cell nuclear antigen.

作者信息

Yamaguchi M, Hayashi Y, Hirose F, Matsuoka S, Moriuchi T, Shiroishi T, Moriwaki K, Matsukage A

机构信息

Laboratory of Cell Biology, Aichi Cancer Center Research Institute, Nagoya, Japan.

出版信息

Nucleic Acids Res. 1991 May 11;19(9):2403-10. doi: 10.1093/nar/19.9.2403.

Abstract

We have isolated clones containing the entire mouse proliferating cell nuclear antigen (PCNA) gene of 3890 bp and flanking sequences using a rat PCNA cDNA as a probe. The mouse gene has 6 exons whose sequences and junction points of exons with introns are extensively homologous to the human gene while sizes and nucleotide sequences of introns are much less conserved than exons. By a transient expression assay of chloramphenicol acetyltransferase, the promoter of this gene is localized within 200 bp upstream of the transcription initiation site. We have also isolated two processed pseudogenes. Homology between the first one (psi PCNA-I) and the exons of the PCNA gene was 76.8% in the region so far sequenced. The second one (psi PCNA-II) consists of a region highly homologous to the entire exons of the PCNA gene, and only 9 out of total 1256 bp are different from the corresponding exon sequence of the gene. The 5'-flanking region of the psi PCNA-II did not function as an active promoter. Surveys in various wild and laboratory mice genomes suggest that the psi PCNA-II was generated through the reverse transcription process of the PCNA mRNA about 5 x 10(5) years ago in the domesticus subspecies of Mus musculus, the house mouse. The psi PCNA-II is tentatively mapped in the chromosome 17 of the C57BL mouse.

摘要

我们使用大鼠增殖细胞核抗原(PCNA)cDNA作为探针,分离出了包含3890 bp完整小鼠PCNA基因及其侧翼序列的克隆。小鼠基因有6个外显子,其序列以及外显子与内含子的连接点与人类基因高度同源,而内含子的大小和核苷酸序列的保守性远低于外显子。通过氯霉素乙酰转移酶的瞬时表达分析,该基因的启动子定位于转录起始位点上游200 bp范围内。我们还分离出了两个加工假基因。到目前为止测序的区域中,第一个假基因(psi PCNA - I)与PCNA基因外显子的同源性为76.8%。第二个假基因(psi PCNA - II)由一个与PCNA基因全部外显子高度同源的区域组成,在总共1256 bp中只有9 bp与该基因相应的外显子序列不同。psi PCNA - II的5'侧翼区域不具有活性启动子的功能。对各种野生和实验室小鼠基因组的调查表明,psi PCNA - II大约在5×10⁵年前通过小家鼠(Mus musculus)的domesticus亚种中PCNA mRNA的逆转录过程产生。psi PCNA - II暂时定位于C57BL小鼠的17号染色体上。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29a1/329449/a4161d0bf6a0/nar00089-0158-a.jpg

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