Huang Chieh-Chen, Chen Meng-Wei, Hsieh Ju-Liang, Lin Wen-Hao, Chen Pei-Chung, Chien Lee-Feng
Department of Life Sciences, National Chung-Hsing University, Tai-Chung 40227, Taiwan.
Appl Microbiol Biotechnol. 2006 Aug;72(1):197-205. doi: 10.1007/s00253-005-0250-0. Epub 2006 Mar 18.
A eukaryotic microalga, Chlorella sp. DT, was transformed with the Bacillus megaterium strain MB1 merA gene, encoding mercuric reductase (MerA), which mediates the reduction of Hg2+ to volatile elemental Hg0. The transformed Chlorella cells were selected first by hygromycin B and then by HgCl2. The existence of merA gene in the genomic DNA of transgenic strains was shown by polymerase chain reaction amplification, while the stable integration of merA into genomic DNA of transgenic strains was confirmed by Southern blot analysis. The ability to remove Hg2+ in merA transgenic strains was higher than that in the wild type. The merA transgenic strains showed higher growth rate and photosynthetic activity than the wild type did in the presence of a toxic concentration of Hg2+. Cultured with Hg2+, the expression level of superoxide dismutase in transgenic strains was lower than that in the wild type, suggesting that the transgenic strains faced a lower level of oxidative stress. All the results indicated that merA gene was successfully integrated into the genome of transgenic strains and functionally expressed to promote the removal of Hg2+.
一种真核微藻,小球藻属DT,用巨大芽孢杆菌菌株MB1的merA基因进行了转化,该基因编码汞还原酶(MerA),可介导Hg2+还原为挥发性元素汞Hg0。首先通过潮霉素B筛选转化的小球藻细胞,然后通过HgCl2筛选。通过聚合酶链反应扩增显示转基因菌株基因组DNA中存在merA基因,而通过Southern印迹分析证实merA稳定整合到转基因菌株的基因组DNA中。merA转基因菌株去除Hg2+的能力高于野生型。在有毒浓度的Hg2+存在下,merA转基因菌株比野生型表现出更高的生长速率和光合活性。用Hg2+培养时,转基因菌株中超氧化物歧化酶的表达水平低于野生型,表明转基因菌株面临较低水平的氧化应激。所有结果表明,merA基因已成功整合到转基因菌株的基因组中并功能性表达,以促进Hg2+的去除。