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当前位置: 首页 > 文章 > NTG诱变筛选赖氨酸缺陷型大肠杆菌 河南农业科学 2015,44 (11) 137-140
Position: Home > Articles > Screening of Lysine Auxotroph Mutant of Escherichia coli by NTG Induced Mutagenesis Journal of Henan Agricultural Sciences 2015,44 (11) 137-140

NTG诱变筛选赖氨酸缺陷型大肠杆菌

作  者：
翟玉洁;姜军坡;王迈;杜国霞;王世英
单  位：
河北农业大学生命科学学院;石家庄市动物卫生监督所
关键词：
亚硝基胍;大肠杆菌;赖氨酸;营养缺陷型;筛选
摘  要：
为了获得产赖氨酸益生菌所需的指示菌株,以大肠杆菌(E.coli)野生型为出发菌株,用0.25 mg/m L的亚硝基胍(NTG)进行诱变,采用青霉素浓缩法淘汰野生型菌株,采用点植对照法在基本培养基(MM)和赖氨酸补充培养基([Lys])上对赖氨酸营养缺陷型(Lys-)E.coli进行初筛,再采用划线对照法在MM和[Lys]上对Lys-疑似菌株进行复检,[Lys]斜面5次传代后,采用划线对照法检测Lys-菌株遗传稳定性。结果表明,0.25 mg/m L NTG处理菌液40 min,对应的致死率为82.4%,经初筛和复检后获得4株Lys-菌株,其中L-3和L-5菌株经5次传代后仍为Lys-菌株,Lys-菌株的检出率为2.70%。
译  名：
Screening of Lysine Auxotroph Mutant of Escherichia coli by NTG Induced Mutagenesis
作  者：
ZHAI Yujie;JIANG Junpo;WANG Mai;DU Guoxia;WANG Shiying;College of Life Science,Agricultural University of Hebei;Animal Quarantine Station of Shijiangzhuang;
关键词：
NTG;;Escherichia coli;;lysine;;auxotroph;;screening
摘  要：
Research was done in order to obtain indicator strains for high efficient screening of probiotic strains which could produce lysine. The wild type strain of Escherichia coli( E. coli) as an original strain was induced for mutagenesis by 0. 25 mg / m L nitrosoguanidine( NTG). The wild type strain was eliminated by the penicillin concentration method. Primary screening of lysine auxotroph mutant( Lys-) strains was done by the control method of point implantation on the minimal medium( MM) and the lysine supplemental medium( [Lys]). Recheck was done for suspected strains by the control method of plate streaking on MM and [Lys]. The genetic stability of Lys-strains were determined by the control method of plate streaking after continuous passaging 5 times on [Lys]inclined medium. Results showed that lethality rate was 82. 4% when broth was induced by NTG for 40 minutes. Four Lys-strains was obtained after primary screening and recheck. Strain L-3 and strain L-5 were still Lys-strains after continuous passaging 5 times on [Lys] inclined medium. And positive rate of Lys-strains was 2. 70%.

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