Dept. of Chemical and Biological Engineering, Colorado State University, Fort Collins, CO 80523, USA.
Biotechnol Prog. 2013 Jan-Feb;29(1):23-30. doi: 10.1002/btpr.1661. Epub 2012 Dec 4.
The cyanobacterium Synechocystis sp. PCC 6803 is a photosynthetic organism capable of efficient harnessing of solar energy while capturing CO(2) 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.
集胞藻 PCC 6803 是一种能够高效利用太阳能并从环境中捕获 CO2 的光合生物。改变其基因组 DNA 的基因修饰方法对于阐明基因功能至关重要，同时也是代谢工程的有用工具。在这项研究中，开发了一种新型的集胞藻基因修饰反选择方法。该方法利用诱导型镍表达 mazF，一种通用的蛋白质合成抑制剂，作为反选择标记。反选择特别有用，因为工程菌株不含任何标记物，这使得进一步的遗传修饰独立于可用的抗生素抗性基因。通过在两种集胞藻变体的几个基因座改变基因，进一步证明了该方法的可用性。
