重组人超氧化物岐化酶基因工程菌稳定性研究  

A Study of the Stability of Recombinant Human Superoxide Dismutase Gene Engineering Bacteria

作　　者：李敏 侯增淼 李晓颖 何越 高恩 杨小琳 赵金礼 

机构地区：[1]陕西慧康生物科技有限责任公司,陕西西安710054

出　　处：《科技创新与应用》2018年第4期11-12,共2页Technology Innovation and Application

摘　　要：目的:研究重组人超氧化物岐化酶毕赤酵母基因工程菌的遗传稳定性。方法:基因工程菌连续传代50次,每10代取样保藏菌种,通过对菌株形态、菌落形态、质粒稳定性、目的基因的PCR鉴定,蛋白表达的SDS-PAGE鉴定来评价工程菌的稳定性。结果:在连续传代50代的过程中,菌株、落形态与原始工程菌相比无明显差异,质粒保有率高,PCR鉴定结果显示,重组载体携带目的基因传至50代未消失,SDS-PAGE结果表明,传至50代蛋白表达量无明显差异。结论:该工程菌有良好的遗传稳定性。Objective： to study the genetic stability of recombinant human superoxide dismutase Pichia pastoris. Methods： the genetically engineered bacteria were subcultured for 50 times, and the preservation strains were sampled every 10 generations. The morphology of the strain, colony morphology, plasmid stability and target gene were identified by PCR. SDS-PAGE analysis of protein expression was used to evaluate the stability of engineering bacteria. Results： there was no significant difference in the strain and colony morphology between the original engineering strain and the original engineering strain in the course of successive subculture for 50 generations. The plasmid retention rate was higher than that of the original engineering strain. The results of PCR identification showed that the plasmid retention rate was high. The result of SDS-PAGE showed that there was no significant difference in the ex-pression of protein from the recombinant vector to the 50th generation. Conclusion： the engineering strain has good genetic stability.

关 键 词：重组人超氧化物岐化酶 毕赤酵母工程菌 遗传稳定性 

分 类 号：Q78[生物学—分子生物学]