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果蝇中QM同源物的分离、定位及特性分析

The isolation, localization and characterization of the QM homolog in Drosophila melanogaster.

作者信息

Nguyen-Yue Y H, Loftus T M, Torok T, Bryant P, Stanbridge E J

机构信息

Department of Microbiology and Molecular Genetics, School of Medicine, University of California, Irvine 92697, USA.

出版信息

DNA Seq. 1997;7(6):337-47. doi: 10.3109/10425179709034053.

Abstract

A fragment of 443 bp was amplified from a lambda ZAPII Drosophila central nervous system (CNS) cDNA library using minimally degenerate primers to very conserved regions of the QM gene. This fragment was used as a probe to screen the lambda ZAPII Drosophila CNS cDNA library. Two clones of the Drosophila QM homolog (pDQM-7A1 and pDQM-2B1), each containing the complete coding region, were isolated. The 5'-UTR of this gene was obtained by RACE PCR and ligated to the coding sequence to produce a the full-length copy of the Drosophila QM homolog (DQM) cDNA. The DQM cDNA measures 746 nucleotides in length and encodes a polypeptide of 218 residues. The amino acid sequence shows 76.1 percent identity with human QM and 69.1 percent identity with QSR1, the yeast homolog of QM. Unlike the human or mouse genome which contains multiple copies of the QM gene, the Drosophila genome has only a single copy as indicated by genomic Southern blot analysis. In situ hybridization confirms the presence of a single copy of DQM in the Drosophila genome and localizes it to the left arm of the third chromosome at the end of region 80A (80A-4).

摘要

使用针对QM基因高度保守区域的最低简并引物,从λZAPII果蝇中枢神经系统(CNS)cDNA文库中扩增出一段443 bp的片段。该片段用作探针筛选λZAPII果蝇CNS cDNA文库。分离出果蝇QM同源物的两个克隆(pDQM - 7A1和pDQM - 2B1),每个克隆都包含完整的编码区。通过RACE PCR获得该基因的5'-UTR,并将其连接到编码序列上,以产生果蝇QM同源物(DQM)cDNA的全长拷贝。DQM cDNA长度为746个核苷酸,编码一个218个残基的多肽。氨基酸序列与人类QM的同一性为76.1%,与QM的酵母同源物QSR1的同一性为69.1%。与包含多个QM基因拷贝的人类或小鼠基因组不同,基因组Southern印迹分析表明果蝇基因组只有一个拷贝。原位杂交证实果蝇基因组中存在单个DQM拷贝,并将其定位到第三条染色体左臂80A区域末端(80A - 4)。

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