MG1655 大肠杆菌-细胞株/菌种-试剂-生物在线
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MG1655 大肠杆菌

MG1655 大肠杆菌

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产品名称: MG1655 大肠杆菌

英文名称: MG1655 大肠杆菌

产品编号: Biovector512362

产品价格: 0

产品产地: Biovector NTCC Inc. USA

品牌商标: Biovector, Addgene, ATCC, Invi

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Order ID

Name

Description

NTCC510822

E.coli MG1655 strain

500uL, Storage:4

Organism:

Escherichia coli (Migula) Castellani and Chalmers

 

Designations:

MG1655

 

Isolation:

derived from existing strain (derived from parent strain W1485 by acridine orange curing of the F plasmid)

 

Antigenic

Properties:

serotype OR:H48:K- 

 

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." (Blattner, et al. 1997). 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 (Jensen 1993). The ilvG- mutation is a frameshift that knocks out acetohydroxy acid synthase II (Lawther, et al. 1982). The rfb-50 mutation is an IS5 insertion that results in the absence of O-antigen synthesis (Liu and Reeves 1994).

MG1655 was derived and named by Mark Guyer from strain W1485, which was derived in Joshua Lederbergs 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 (Bachmann, B., pp. 2460-2488 in Neidhardt et al.1996, Escherichia coli and Salmonella: Cellular and Molecular Biology, ASM Press).

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 Neidhardts 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 Neidhardts medium because it shows a sharper transition to stationary phase.

 

Recovery

  1. Obtain an LB agar plate with the appropriate antibiotic.
  2. Using a sterile pipette tip, touch the bacteria growing within the punctured area of the stab culture. (A sterilized wire loop or sterile toothpick can be used in place of a sterile pipette tip.)
  3. Run this tip lightly over a section of the plate, as shown in the figure, to create streak #1.
  4. Using another sterile pipette tip, pass through streak #1 and spread the bacteria over a second section of the plate, to create streak #2.
  5. Using a third sterile pipette tip, pass through streak #2 and spread the bacteria over the last section of the plate, to create streak #3.
  6. Grow overnight in a 37 o C incubator (unless a different growth temperature is indicated on the plasmid datasheet).
  7. In the morning, single colonies should be visible. If the bacterial growth is too dense, re-streak onto a new agar plate to obtain single colonies.