Fujimura H, Sakuma Y, Amann E
Laboratory of Drug Creation Research, Hoechst Japan Limited, Kawagoe.
J Appl Bacteriol. 1994 Dec;77(6):689-93. doi: 10.1111/j.1365-2672.1994.tb02820.x.
A genetically-engineered strain of Saccharomyces cerevisiae employed for the industrial production of the human coagulation Factor XIIIa (rhFXIIIa) was used for a survival study under simulated environmental conditions. The homologous strain devoid of the recombinant plasmid and the homologous strain bearing the 2 microns-based vector plasmid without the rhFXIIIa-encoding DNA insert were compared. The strains were introduced into natural soil/water suspension, into soil/medium suspension and into waste water. After intervals, samples of cell suspensions were taken and viable cell numbers were determined by plating on antibiotic-containing medium. In addition, a non-radioactive technique involving enhanced chemiluminescence was employed to detect plasmid-bearing yeast cells. The rhFXIIIa expression plasmid showed a high stability during the simulated environmental condition. No differences in survival rates, however, could be detected for the plasmid-bearing and plasmid-less strains under the three conditions tested, suggesting that the presence of plasmid does not confer selective advantages on the survival of the yeast cells. It is concluded that, even after accidental release of the engineered yeast cells into the environment, elimination rates would be comparable to those for non-recombinant yeast strains.
一种用于工业生产人凝血因子XIIIa(rhFXIIIa)的基因工程酿酒酵母菌株被用于模拟环境条件下的存活研究。将不含重组质粒的同源菌株和携带不含rhFXIIIa编码DNA插入片段的基于2微米载体质粒的同源菌株进行比较。将这些菌株引入天然土壤/水悬浮液、土壤/培养基悬浮液和废水中。经过一定时间间隔后,采集细胞悬浮液样本,并通过在含抗生素培养基上平板接种来测定活细胞数量。此外,采用一种涉及增强化学发光的非放射性技术来检测携带质粒的酵母细胞。rhFXIIIa表达质粒在模拟环境条件下显示出高稳定性。然而,在测试的三种条件下,携带质粒和无质粒的菌株在存活率上没有差异,这表明质粒的存在并未赋予酵母细胞存活的选择优势。得出的结论是,即使工程酵母细胞意外释放到环境中,消除率也将与非重组酵母菌株相当。