Kang Nam Kyu, Choi Gang-Guk, Kim Eun Kyung, Shin Sung-Eun, Jeon Seungjib, Park Min S, Jeong Ki Jun, Jeong Byeong-Ryool, Chang Yong Keun, Yang Ji-Won, Lee Bongsoo
Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Advanced Biomass R&D Center, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Biotechnol Rep (Amst). 2015 Aug 22;8:10-15. doi: 10.1016/j.btre.2015.08.004. eCollection 2015 Dec.
Oleaginous microalgae of the genus are considered excellent candidates for biofuels and value-added products owing to their high biomass productivity and lipid content. Here, we report the first overexpression and detection of a heterologous sfCherry fluorescent protein in in order to develop a transformation toolbox for future genetic improvements. Particle bombardment was employed for transformation, and expression of Sh under the control of TUB and UEP promoters, cloned from , was used to confer resistance to Zeocin antibiotics, resulting in 5.9 and 4.7 transformants per 10 cells, respectively. Stable integration of the markers into the genome was confirmed using a restriction enzyme site-directed amplification (RESDA) PCR. The expression of sfCherry fluorescent protein was confirmed by Western blot analysis and confocal microscopy. These results suggest new possibilities of efficient genetic engineering of for the production of biofuels and other biochemicals.
由于其高生物量生产力和脂质含量,该属的产油微藻被认为是生物燃料和增值产品的优秀候选者。在此,我们报告了首次在[具体物种]中过表达和检测异源sfCherry荧光蛋白,以便为未来的基因改良开发一个转化工具箱。采用粒子轰击进行转化,并使用从[具体物种]克隆的TUB和UEP启动子控制下的Sh表达来赋予对博来霉素抗生素的抗性,每10个细胞分别产生5.9个和4.7个转化体。使用限制性酶切位点定向扩增(RESDA)PCR确认标记物稳定整合到基因组中。通过蛋白质免疫印迹分析和共聚焦显微镜确认sfCherry荧光蛋白的表达。这些结果表明了对[具体物种]进行高效基因工程以生产生物燃料和其他生化物质的新可能性。