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盘基网柄菌核糖体蛋白基因的鉴定与表征

Identification and characterization of a Dictyostelium discoideum ribosomal protein gene.

作者信息

Szymkowski D E, Deering R A

机构信息

Department of Molecular and Cell Biology, Pennsylvania State University, University Park, PA 16802.

出版信息

Nucleic Acids Res. 1990 Aug 25;18(16):4695-701. doi: 10.1093/nar/18.16.4695.

Abstract

We have identified a developmentally repressed large-subunit ribosomal protein gene of Dictyostelium discoideum based on sequence similarity to other ribosomal proteins. Protein rpl7 is homologous to large subunit ribosomal proteins from the rat and possibly to Mycoplasma capricolum and Escherichia coli, but is not similar to three sequenced ribosomal proteins in Dictyostelium. The rpl7 gene is present at one copy per genome, as are six other cloned Dictyostelium ribosomal proteins. Restriction fragment length polymorphisms exist for ribosomal protein genes rpl7, rp1024, and rp110 in strain HU182; most Dictyostelium ribosomal protein genes examined are linked no closer than 30-100 kb to each other in the genome. Dictyostelium ribosomal proteins are known to be developmentally regulated, and levels of rpl7 transcript gradually decrease during the 24-hour development cycle. This drop correlates with that of rp1024, indicating these and other ribosomal protein genes may be coordinately regulated. To determine the cellular location of the protein, we raised antibodies to an rpl7-derived branched synthetic peptide. These antibodies cross-reacted with one protein of the expected size in a ribosomal protein fraction of Dictyostelium, indicating that the product of gene rpl7 is localized in the ribosome.

摘要

基于与其他核糖体蛋白的序列相似性,我们鉴定出了盘基网柄菌中一个在发育过程中受到抑制的大亚基核糖体蛋白基因。蛋白质rpl7与大鼠的大亚基核糖体蛋白同源,可能也与山羊支原体和大肠杆菌的大亚基核糖体蛋白同源,但与盘基网柄菌中已测序的三种核糖体蛋白不相似。rpl7基因在每个基因组中只有一个拷贝,其他六个已克隆的盘基网柄菌核糖体蛋白基因也是如此。在HU182菌株中,核糖体蛋白基因rpl7、rp1024和rp110存在限制性片段长度多态性;在基因组中,大多数被检测的盘基网柄菌核糖体蛋白基因彼此之间的连锁距离不小于30 - 100 kb。已知盘基网柄菌核糖体蛋白受到发育调控,在24小时的发育周期中,rpl7转录本水平逐渐降低。这种下降与rp1024的下降相关,表明这些核糖体蛋白基因和其他核糖体蛋白基因可能受到协同调控。为了确定该蛋白的细胞定位,我们制备了针对rpl7衍生的分支合成肽的抗体。这些抗体与盘基网柄菌核糖体蛋白组分中预期大小的一种蛋白发生交叉反应,表明rpl7基因的产物定位于核糖体中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b64b/331922/ceed947c09dd/nar00200-0058-a.jpg

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