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用鼠伤寒沙门氏菌菌株SV3进行的L-阿拉伯糖抗性试验可筛选多个阿拉伯糖基因的正向突变。

The L-arabinose-resistance test with Salmonella typhimurium strain SV3 selects forward mutations at several ara genes.

作者信息

Pueyo C, Lopez-Barea J

出版信息

Mutat Res. 1979 Aug;64(4):249-58. doi: 10.1016/0165-1161(79)90094-3.

Abstract

A new assay has been described for mutagenicity testing using an L-arabinose-sensitive strain of Salmonella typhimurium. The test strain SV3 and several L-arabinose-resistant mutants selected therefrom are characterized in the present study by 3 different criteria: inhibition of growth by L-arabinose, accumulation of keto-sugars, and activities of the enzymes involved in L-arabinose catabolism. Strain SV3 (ara-531) shows high levels of inducible L-arabinose isomerase (EC 5.3.1.4) and L-ribulokinase (EC 2.7.1.16) activities, but is deficient in L-ribulose-5-phosphate 4-epimerase (EC 5.1.3.4), the enzyme encoded in Escherichia coli by gene D in the araBAD operon. Addition of L-arabinose to SV3 growing in glycerol or casamino acids stops growth. D-Glucose only partially reverses this inhibition. Reversion of the ara-531 mutation restores different levels of epimerase activity and resistance to L-arabinose. However, the great majority of the L-arabinose-resistant mutants do not utilize L-arabinose. The physiological and enzymatic properties of these L-arabinose non-utilizing mutants suggest that L-arabinose resistance is due to forward mutations in at least 3 other genes, araA, araB and araC, blocking steps prior to L-ribulose 5-phosphate accumulation.

摘要

已描述了一种新的用于致突变性测试的方法,该方法使用鼠伤寒沙门氏菌的一种对L-阿拉伯糖敏感的菌株。在本研究中,通过3种不同标准对测试菌株SV3及其从中筛选出的几种L-阿拉伯糖抗性突变体进行了表征:L-阿拉伯糖对生长的抑制作用、酮糖的积累以及参与L-阿拉伯糖分解代谢的酶的活性。菌株SV3(ara-531)显示出高水平的可诱导L-阿拉伯糖异构酶(EC 5.3.1.4)和L-核糖激酶(EC 2.7.1.16)活性,但缺乏L-核糖ulose-5-磷酸4-表异构酶(EC 5.1.3.4),该酶在大肠杆菌中由araBAD操纵子中的基因D编码。向在甘油或酪蛋白氨基酸中生长的SV3添加L-阿拉伯糖会停止生长。D-葡萄糖只能部分逆转这种抑制作用。ara-531突变的回复会恢复不同水平的表异构酶活性和对L-阿拉伯糖的抗性。然而,绝大多数L-阿拉伯糖抗性突变体不利用L-阿拉伯糖。这些不利用L-阿拉伯糖的突变体的生理和酶学特性表明,L-阿拉伯糖抗性是由于至少其他3个基因araA、araB和araC中的正向突变,这些突变阻断了L-核糖ulose 5-磷酸积累之前的步骤。

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