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对照和热激处理的玉米幼苗组织中不同多聚泛素基因的表征、染色体定位及表达

Characterization, chromosomal mapping, and expression of different polyubiquitin genes in tissues from control and heat-shocked maize seedlings.

作者信息

Liu L, Maillet D S, Frappier J R, Walden D B, Atkinson B G

机构信息

Department of Zoology, University of Western Ontario, London, Canada.

出版信息

Biochem Cell Biol. 1995 Jan-Feb;73(1-2):19-30. doi: 10.1139/o95-003.

Abstract

Polyubiquitin transcripts accumulate in plant and animal cells following a heat shock. Most species have a few to several polyubiquitin genes; within a species, the genes may differ in nucleotide (nt) sequence and (or) the number of 228-nt repeats encoding the ubiquitin monomer. This study examines three maize (inbred Oh43) polyubiquitin genes. Two of the genes, MubG9 and MubG5, possess five repeats; the third, MubG1 possesses seven repeats. Sequence analyses of the genomic clones, MubG9 and MubG1 and a cDNA clone, MubG5, reveal that they differ primarily from each other in their nt sequences 5' and 3' to their open reading frames. MubG1 contains a 1004-base pair (bp) intron in its 5' untranslated region. Using gene-specific probes, we show that the amount of polyribosome-associated mRNA transcripts from MubG9 isolated from 2- and 5-day old plumules and radicles is unchanged by heat shock. While the amount of transcript from MubG1 and MubG5 on the polyribosomes in plumules and radicles of 2-day-old seedlings is also unchanged by heat shock, the levels of these transcripts are elevated considerably in similar tissues from heat-shocked 5-day-old seedlings. Similar or identical gene-specific probes were employed to map the genes using the recombinant inbred method. MubG9 maps to chromosome 4L position 186; MubG1 maps to 5L104 and MubG5 to 4L188. The opportunity to use gene-specific probes extends the evidence for distinct modulation (time and tissue) of polyubiquitin gene expression in maize and provides the basis for locus assignment within the genome.

摘要

热激后,多聚泛素转录本在植物和动物细胞中积累。大多数物种有几个到几个多聚泛素基因;在一个物种内,这些基因在核苷酸(nt)序列和(或)编码泛素单体的228 nt重复序列的数量上可能有所不同。本研究检测了三个玉米(自交系Oh43)多聚泛素基因。其中两个基因,MubG9和MubG5,有五个重复序列;第三个基因MubG1有七个重复序列。对基因组克隆MubG9和MubG1以及cDNA克隆MubG5的序列分析表明,它们彼此之间的主要差异在于其开放阅读框5'和3'端的nt序列。MubG1在其5'非翻译区含有一个1004碱基对(bp)的内含子。使用基因特异性探针,我们发现从2日龄和5日龄的胚芽和胚根中分离出的MubG9的多核糖体相关mRNA转录本的量不受热激影响。虽然2日龄幼苗的胚芽和胚根中多核糖体上MubG1和MubG5的转录本量也不受热激影响,但在热激处理的5日龄幼苗的类似组织中,这些转录本的水平显著升高。使用相似或相同的基因特异性探针,采用重组自交法对这些基因进行定位。MubG9定位于第4L染色体186位置;MubG1定位于5L104,MubG5定位于4L188。使用基因特异性探针的机会扩展了玉米中多聚泛素基因表达独特调控(时间和组织)的证据,并为基因组内基因座的定位提供了基础。

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