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海草无轴草Cymodocea nodosa的钾转运蛋白CnHAK1和CnHAK2在细菌中的分子克隆及功能表达

Molecular cloning and functional expression in bacteria of the potassium transporters CnHAK1 and CnHAK2 of the seagrass Cymodocea nodosa.

作者信息

Garciadeblas Blanca, Benito Begoña, Rodríguez-Navarro Alonso

机构信息

Departamento de Biotecnología, Escuela Técnica Superior de Ingenieros Agrónomos, Universidad Politécnica de Madrid, Spain.

出版信息

Plant Mol Biol. 2002 Nov;50(4-5):623-33. doi: 10.1023/a:1019951023362.

Abstract

The cDNAs CnHAK1 and CnHAK2, encoding K+ transporters, were amplified from the leaves of the seagrass Cymodocea nodosa. None of these transporters suppressed the K+ deficiency of a Saccharomyces cerevisiae mutant, but both suppressed the equivalent defect of an Escherichia coli mutant. Overexpression of the transporter CnHAKI, but not CnHAK2, mediated very rapid K+ or Rb+ influxes in the E. coli mutant. The concentration dependence of these influxes demonstrated that CnHAK1 is a low-affinity K+ transporter, which does not discriminate between K+ and Rb+. CnHAK1 expressed in E. coli worked in reverse when the external K+ concentrations were low, and we established the condition of a simple functional test of K+ loss for transporters of the Kup-HAK family. In comparison with its homologue barley transporter HvHAK2, CnHAKI was insensitive to Na+.

摘要

编码钾离子转运蛋白的cDNA CnHAK1和CnHAK2是从海草波喜荡草的叶片中扩增得到的。这些转运蛋白均不能抑制酿酒酵母突变体的钾离子缺乏,但二者都能抑制大肠杆菌突变体的同等缺陷。转运蛋白CnHAKI(而非CnHAK2)的过表达介导了大肠杆菌突变体中非常快速的钾离子或铷离子内流。这些内流的浓度依赖性表明CnHAK1是一种低亲和力的钾离子转运蛋白,它不能区分钾离子和铷离子。当外部钾离子浓度较低时,在大肠杆菌中表达的CnHAK1会反向工作,并且我们建立了一种针对Kup-HAK家族转运蛋白的简单钾离子流失功能测试条件。与其同源物大麦转运蛋白HvHAK2相比,CnHAKI对钠离子不敏感。

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