[4]膜蛋白RHOGL009301参与红球菌中苯甲酸盐运输的生理功能[j]。

微生物学报 Pub Date : 2017-04-04
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引用次数: 0

摘要

目的:研究红球菌R04中膜蛋白RHOGL009301的生理功能及突变菌株的代谢特性,以确定膜蛋白的生理功能与苯甲酸盐转运的关系。方法:将RHOGL009301基因与绿色荧光蛋白基因融合在红红球菌中表达,用Delta Vision观察RHOGL009301基因的位置。通过同源重组敲除RHOGL009301基因,比较了野生菌株和缺陷菌株在不同碳源下的生长情况。采用高效液相色谱法测定了野生菌株和缺陷菌株在联苯和苯甲酸酯上生长时的内外代谢产物,并分析了不同生长条件下代谢产物浓度的变化。结果:一个包含RHOGL009301基因和绿色荧光蛋白基因的融合基因在红红球菌中共表达,并定位在细胞膜上。获得RHOGL009301基因缺失株R04ΔMP。在联苯和苯甲酸酯培养中,缺陷菌株的生物量显著降低,生长速度减慢。HPLC分析显示RHOGL009301基因的缺失抑制了苯甲酸盐的转运。结论:RHOGL009301膜蛋白是参与苯甲酸盐代谢和转运的蛋白之一。根据序列同源性分析,该膜蛋白是一种新型的苯甲酸盐转运蛋白。
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[Physiological function of membrane protein RHOGL009301 involved in transport of benzoate in Rhodococcus sp. R04].

Objective: The physiological function of membrane protein RHOGL009301 in Rhodococcus sp. R04 and the metabolic properties of the mutant strain were studied to determine the relationship between the physiological function of the membrane protein and the transport of benzoate.

Methods: The RHOGL009301 gene and the green fluorescent protein gene were fused for expressing in Rhodococcus erythropolis, and the location of RHOGL009301 was observed by Delta Vision. The RHOGL009301 gene was knocked out by homologous recombination, and the growth of wild strain and deficient strain in different carbon sources were compared. The internal and external metabolites of the wild strain and the deficient strain when grown on biphenyl and benzoate were measured by HPLC, and the changes of metabolite concentration in different growth conditions were analyzed.

Results: A fusion gene that contained RHOGL009301 gene and the green fluorescent protein gene was co-expressed in Rhodococcus erythropolis and localized on the cell membrane. The deficient strain R04ΔMP of RHOGL009301 gene was obtained. The biomass of the deficient strain was significantly reduced in biphenyl and benzoate culture, and its growth rate was slowed down. HPLC analysis showed that the deletion of RHOGL009301 gene inhibited the transport of benzoate.

Conclusion: RHOGL009301 membrane protein is one of the proteins involved in metabolism and transport of benzoate. Based on sequence homology analysis, we can conclude that the membrane protein is a novel benzoate transport protein.

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期刊介绍: Acta Microbiologica Sinica(AMS) is a peer-reviewed monthly (one volume per year)international journal,founded in 1953.It covers a wide range of topics in the areas of general and applied microbiology.The journal publishes original papers,reviews in microbiological science,and short communications describing unusual observations. Acta Microbiologica Sinica has been indexed in Index Copernicus (IC),Chemical Abstract (CA),Excerpt Medica Database (EMBASE),AJ of Viniti (Russia),Biological Abstracts (BA),Chinese Science Citation Database (CSCD),China National Knowledge Infrastructure(CNKI),Institute of Scientific and Technical Information of China(ISTIC),Chinese Journal Citation Report(CJCR),Chinese Biological Abstracts,Chinese Pharmaceutical Abstracts,Chinese Medical Abstracts and Chinese Science Abstracts.
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