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通过呋喃妥因处理高效筛选集胞藻PCC 6803光合作用缺陷突变体条件的建立。

The establishment of conditions to efficiently screen photosynthesis-deficient mutants of Synechocystis sp. PCC 6803 by nitrofurantoin treatment.

作者信息

Yamasato A, Satoh K

机构信息

Graduate School of Natural Science and Technology, Okayama University, Okayama, 700-8530 Japan.

出版信息

Plant Cell Physiol. 2001 Apr;42(4):414-8. doi: 10.1093/pcp/pce051.

Abstract

A method based on the susceptibility of photosynthetic organisms to nitrofurantoin under illumination was established to screen mutants of Synechocystis sp. PCC 6803 deficient in the function of photosystem II, which were created by random PCR mutagenesis targeted to the psbAII gene coding for the D1 protein of the photosystem II reaction center. In this method, cyanobacterial colonies on a nitrocellulose membrane on a BG11 agar plate were treated with nitrofurantoin at 1.0 mM under white light at 40 microE x m(-2 ) x s(-1) for 2 h, and then kept under normal conditions without nitrofurantoin so that surviving cells could grow. This method was also shown to be useful for screening mutants deficient in the function of photosystem I.

摘要

建立了一种基于光合生物在光照下对呋喃妥因敏感性的方法,用于筛选集胞藻6803(Synechocystis sp. PCC 6803)中光系统II功能缺陷的突变体,这些突变体是通过针对编码光系统II反应中心D1蛋白的psbAII基因进行随机PCR诱变产生的。在该方法中,将BG11琼脂平板上硝酸纤维素膜上的蓝细菌菌落置于40 μE×m⁻²×s⁻¹的白光下,用1.0 mM的呋喃妥因处理2小时,然后在无呋喃妥因的正常条件下培养,使存活的细胞能够生长。该方法也被证明可用于筛选光系统I功能缺陷的突变体。

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