食品工业科技2017，Vol.38Issue(19)：114-117,124,5.DOI:10.13386/j.issn1002-0306.2017.19.022
生长分化因子(BMP11)重组质粒在工程菌株中的稳定性研究
Study on the stability of growth differentiation factor (BMP11) recombinant plasmid in Escherichia coli
摘要
Abstract
To investigate the hereditary stability of recombinant plasmid (pET-BMP11)in host cell (Escherichiacoli BL21 (DE3)).The recombinant E.coli was subcultured for 100 generation,samples were collected every 20 generations.Bacterial morphology,bacteria growth,plasmid stability,the target protein expressing level were analyzed.The Xho Ⅰ and Nde Ⅰ double enzyme validation and determination of target sequence were performed on the origin,20th,60th,100th generation strain.The results showed that the growth characteristics and the morphology of the cultures did not show differences among the generations,in the absence of kanamycin.The plasmid stability and protein expression of 40th generation strain decreased significantly,and the plasmid retention rate of 100th generations was only 56％.In the presence of kanamycin,the 80th generation strain still had a high level of protein expression.The restriction enzyme map and target gene sequence of 100th were consistent with the original ones,and the rate of plasmid retention was up to 96％.These results showed that the use of recombinant E.coli to produce BMP11 was feasible,the addition of kanamycin in the medium kept the recombinant plasmid stable during passage,this study could provide a theoretical basis for the industrial production of BMP11 and the development of new anti-aging foods.关键词
生长分化因子11/重组质粒/工程菌株/稳定性Key words
growth differentiation factor 11/recombinant plasmid/engineering strain/stability分类
轻工纺织引用本文复制引用
郝丹,尤倩倩,郝伟,梁晓琳,袁靖琳,李全阳..生长分化因子(BMP11)重组质粒在工程菌株中的稳定性研究[J].食品工业科技,2017,38(19):114-117,124,5.基金项目
国家自然科学基金面上项目(31371762). （31371762）
