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蚜虫内共生菌——蚜虫内共生菌(Buchnera aphidicola)的核糖体蛋白S1(RpsA):基因特征与产物检测

Ribosomal protein S1 (RpsA) of Buchnera aphidicola, the endosymbiont of aphids: characterization of the gene and detection of the product.

作者信息

Clark M A, Baumann L, Baumann P, Rouhbakhsh D

机构信息

Microbiology Section, University of California, Davis 95616-8665, USA.

出版信息

Curr Microbiol. 1996 Feb;32(2):89-94. doi: 10.1007/s002849900016.

Abstract

Buchnera aphidicola is a prokaryotic endosymbiont found in specialized cells of the aphid Schizaphis graminum. Many of the previously cloned B. aphidicola genes are preceded by a poor ribosome-binding site. Ribosomal protein S1 (RpsA) allows the translation of messenger RNAs that lack or have a poor ribosome binding site. We have cloned and sequenced a 4.5-kilobase (kb) B. aphidicola DNA fragment containing four open reading frames corresponding to aroA-rpsA-himD-tpiA. The deduced amino acid sequence of B. aphidicola RpsA was 75% identical to that of the Escherichia coli protein. The major difference was in the number of basic amino acids, which were present in higher numbers in B. aphidicola RpsA. Antiserum to E. coli RpsA was prepared and used to detect B. aphidicola RpsA in cell-free extracts of aphids. During the first 12 days of aphid growth there is a slight decrease in the amount of RpsA per unit of aphid weight. The three additional genes found on the 4.5-kb DNA fragment encoded for proteins involved in aromatic amino acid biosynthesis (aroA), DNA bending (himD), and carbohydrate metabolism (tpiA). The presence of these genes in B. aphidicola is additional evidence of its similarity to free-living bacteria.

摘要

蚜虫内共生菌是一种原核内共生体,存在于麦二叉蚜的特化细胞中。许多先前克隆的蚜虫内共生菌基因之前都有一个较差的核糖体结合位点。核糖体蛋白S1(RpsA)能使缺乏或具有较差核糖体结合位点的信使核糖核酸进行翻译。我们克隆并测序了一段4.5千碱基(kb)的蚜虫内共生菌DNA片段,该片段包含四个开放阅读框,分别对应aroA - rpsA - himD - tpiA。蚜虫内共生菌RpsA推导的氨基酸序列与大肠杆菌蛋白的氨基酸序列有75%的同一性。主要差异在于碱性氨基酸的数量,蚜虫内共生菌RpsA中的碱性氨基酸数量更多。制备了针对大肠杆菌RpsA的抗血清,并用于检测蚜虫无细胞提取物中的蚜虫内共生菌RpsA。在蚜虫生长的前12天,每单位蚜虫体重的RpsA量略有下降。在4.5 kb DNA片段上发现的另外三个基因编码参与芳香族氨基酸生物合成(aroA)、DNA弯曲(himD)和碳水化合物代谢(tpiA)的蛋白质。这些基因在蚜虫内共生菌中的存在进一步证明了它与自由生活细菌的相似性。

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