Oh Eun-Taex, So Jae-Seong, Kim Byung-Hyuk, Kim Jong-Sul, Koh Sung-Cheol
Department of Biological Engineering, Center for Advanced Bioseparation Technology, Inha University, Incheon 402-751, Korea.
J Microbiol. 2004 Sep;42(3):243-7.
Sphingomonas chlorophenolica ATCC39723 was successfully labeled with the gfp (green fluorescent protein) gene inserted into the pcpB gene by homologous recombination. As the gfp recombinant was easily distinguished from other indigenous organisms, the population of gfp recombinant was monitored after being released into the soil microcosms. Their population density dropped from 10(8) to 10(6) (cfu/ml) in the non-sterilized soil microcosms during the first 6 days. Moreover, the gfp recombinant was not detected even at lower dilution rates after a certain time period. The recombinant, however, survived for at least 28 days in the sterilized soil microcosms. Although the gfp recombinant did not degrade pentachlorophenol (PCP), this experiment showed the possibility of using gfp as a monitoring reporter system for S. chlorophenolica ATCC39723 and potentially other species of Sphingomonas.
通过同源重组将绿色荧光蛋白(gfp)基因插入到嗜氯鞘氨醇单胞菌(Sphingomonas chlorophenolica)ATCC39723的对氯苯酚降解基因(pcpB)中,成功地对该菌进行了标记。由于绿色荧光蛋白重组体很容易与其他本土生物区分开来,因此在将其释放到土壤微观生态系统后,对绿色荧光蛋白重组体的数量进行了监测。在未灭菌的土壤微观生态系统中,其种群密度在最初6天内从10⁸下降到10⁶(cfu/ml)。此外,在一段时间后,即使在较低稀释率下也检测不到绿色荧光蛋白重组体。然而,该重组体在灭菌的土壤微观生态系统中存活了至少28天。尽管绿色荧光蛋白重组体不能降解五氯苯酚(PCP),但该实验表明了使用绿色荧光蛋白作为嗜氯鞘氨醇单胞菌ATCC39723以及潜在的其他鞘氨醇单胞菌属物种的监测报告系统的可能性。