A novel counter-selection method for markerless genetic modification in Synechocystis sp PCC 6803
A novel counter-selection method for markerless genetic modification in Synechocystis sp PCC 6803
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DOI：
10.1002/btpr.1661
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发表时间：
2013-01-01
影响因子：
2.9
通讯作者：
Peebles, Christie A. M.
中科院分区：
文献类型：
--
作者：
Cheah, Yi Ern;Albers, Stevan C.;Peebles, Christie A. M.
蓝细菌集胞藻PCC 6803是一种光合生物，能够有效利用太阳能，同时从环境中捕获CO2。改变其基因组DNA的方法对于阐明基因功能是必不可少的，并且是代谢工程的有用工具。在这项研究中，一个新的反选择方法的遗传变异的集胞藻。该方法利用镍诱导的mazF（一种通用蛋白质合成抑制剂）表达作为反选择标记。反选择是特别有用的，因为工程菌株不含任何标记物，所述标记物使进一步的遗传修饰不依赖于可用的抗生素抗性基因。通过改变集胞藻两个变种中几个位点的基因，进一步证明了该方法的可用性。(c)2012年美国化学工程师学会生物技术。Prog. 2013年29日、23日至30日
The cyanobacterium Synechocystis sp. PCC 6803 is a photosynthetic organism capable of efficient harnessing of solar energy while capturing CO2 from the environment. Methods to genetically alter its genomic DNA are essential for elucidating gene functions and are useful tools for metabolic engineering. In this study, a novel counter-selection method for the genetic alteration of Synechocystis was developed. This method utilizes the nickel inducible expression of mazF, a general protein synthesis inhibitor, as a counter-selection marker. Counter-selection is particularly useful because the engineered strain is free of any markers which make further genetic modification independent of available antibiotic resistance genes. The usability of this method was further demonstrated by altering genes at several loci in two variants of Synechocystis. (c) 2012 American Institute of Chemical Engineers Biotechnol. Prog. 29;23-30, 2013