E. coli K-12 MG1655 NTCC®大肠杆菌专利菌株-BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

E. coli K-12 MG1655 NTCC®大肠杆菌专利菌株-BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

E. coli K-12 MG1655

Description

Genotype: F- lambda- ilvG- rfb-50 rph-1

Serotype: OR:H48:K-

This strain was sequenced by the Blattner laboratory because it approximates wild-type E. coli and "has been maintained as a laboratory strain with minimal genetic manipulation, having only been cured of the temperate bacteriophage lambda and F plasmid by means of ultraviolet light and acridine orange, respectively." [1]. The mutations listed in the genotype are present in most K-12 strains and were probably acquired early in the history of the laboratory strain. A frameshift at the end of rph results in decreased pyrE expression and a mild pyrimidine starvation, such that the strain grows 10 to 15% more slowly in pyrimidine-free medium than in medium containing uracil [2]. The ilvG- mutation is a frameshift that knocks out acetohydroxy acid synthase II [3]. The rfb-50 mutation is an IS5 insertion that results in the absence of O-antigen synthesis [4].

MG1655 was derived and named by Mark Guyer from strain W1485, which was derived in Joshua Lederberg's lab from a stab-culture descendant of the original K-12 isolate. This original E. coli strain K-12 was obtained from a stool sample of a diphtheria patient in Palo Alto, CA in 1922 [5].

1. Blattner, et al (1997) The complete genome sequence of Escherichia coli K-12. Science 277(5331), 1453-1462. PMID: 9278503

2. Jensen (1993) The Escherichia coli K-12 "wild types" W3110 and MG1655 have an rph frameshift mutation that leads to pyrimidine starvation due to low pyrE expression levels. J Bacteriol 175(11), 3401-3407. PMID: 8501045

3. Lawther, et al. (1982) DNA sequence fine-structure analysis of ilvG (IlvG+) mutations of Escherichia coli K-12. J Bacteriol 149(1), 294-298. PMID: 7033211

4. Liu and Reeves (1994) Escherichia coli K12 regains its O antigen. Microbiology 140(Pt 1), 49-57. PMID: 7512872

5. Bachmann (1996) Derivations and Genotypes of Some Mutant Derivatives of Escherichia coli K-12. p2460-2488 in Neidhardt (ed), Escherichia coli and Salmonella, 2nd Edition ASM Press, Washington DC

Media and growth curves

MG1655 grows on LB and M9 minimal medium (+ Glucose + 1ug/ml thiamine). In doing experiments with microarrays, we sought a medium that was both defined and reproducible (unlike LB), yet supported fast growth rates. We now use Neidhardt's MOPS-based rich defined medium (MOPS-RDM). We also sought a commercially available rich defined medium, and finding none tried to grow MG1655 on a medium sold for mammalian cell culture. It grows quite well, but we decided to stick with Neidhardt's medium because it shows a sharper transition to stationary phase.

Recipe for MOPS Rich Defined Medium and MOPS Minimal Medium

Growth curve for MG1655 on MOPS Minimal Medium

Information and growth curve for a commercially available rich defined medium (MEM alpha w/o phenol red)

Growth curves for MG1655 on M9 Minimal Medium

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