Small membrane proteins found by comparative genomics and ribosome binding site models.

Small membrane proteins found by comparative genomics and ribosome binding site models.
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通过比较基因组学和核糖体结合位点模型发现的小膜蛋白。

DOI:
10.1111/j.1365-2958.2008.06495.x
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发表时间:
2008-12
影响因子:
3.6
通讯作者:
Rudd KE
Rudd KE
中科院分区:
生物学2区
文献类型:
--
作者:
Hemm MR;Paul BJ;Schneider TD;Storz G;Rudd KE

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编码最小蛋白质的基因的正确注释是基因组注释的最大挑战之一,也许更重要的是,很少有注释的短ORF被证实对应于合成的蛋白质。我们使用序列保守和核糖体结合位点模型来预测编码小蛋白的基因,定义为具有16-50个氨基酸,在大肠杆菌基因组的基因间区域。我们测试了这些预测的以及先前注释的基因的表达,通过整合顺序肽亲和标签直接上游的终止密码子的染色体上,并测定合成使用免疫印迹分析。该方法证实了20个先前注释的和18个新发现的16-50个氨基酸的蛋白质被合成。我们总结了这些小蛋白质的性质,显着超过一半的蛋白质被预测为单跨膜蛋白质,其中9个,我们显示co-fractionary与细胞膜。
The correct annotation of genes encoding the smallest proteins is one of the biggest challenges of genome annotation, and perhaps more importantly, few annotated short ORFs have been confirmed to correspond to synthesized proteins. We used sequence conservation and ribosome binding site models to predict genes encoding small proteins, defined as having 16-50 amino acids, in the intergenic regions of the Escherichia coli genome. We tested expression of these predicted as well as previously-annotated genes by integrating the sequential peptide affinity tag directly upstream of the stop codon on the chromosome and assaying for synthesis using immunoblot assays. This approach confirmed that 20 previously-annotated and 18 newly-discovered proteins of 16-50 amino acids are synthesized. We summarize the properties of these small proteins; remarkably more than half of the proteins are predicted to be single transmembrane proteins, nine of which we show co-fractionate with cell membranes.
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