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Team:Kyoto/Project/Goal B
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==Goal B: Characterization of λ Lysis cassette== | ==Goal B: Characterization of λ Lysis cassette== | ||
===Introduction=== | ===Introduction=== | ||
| - | λ Lysis cassette is a gene that causes cell lysis.To characterize the lytic activity of λ Lysis cassette quantitatively, we regulate the gene expression of λ Lysis cassette by a lactose promoter, R0011, which we characterized quantitatively by RPU in GoalA. To repress the basal expression of λ Lysis cassette without IPTG as possible, we used pSB4K5, a low copy vector, as a plasmid backbone and used KRX as a host cell. Needless to say, we characterized R0011 in the same experimental condition.For more detailed explanation of the characterization of R0011, go | + | λ Lysis cassette is a gene that causes cell lysis.To characterize the lytic activity of λ Lysis cassette quantitatively, we regulate the gene expression of λ Lysis cassette by a lactose promoter, R0011, which we characterized quantitatively by RPU in GoalA. To repress the basal expression of λ Lysis cassette without IPTG as possible, we used pSB4K5, a low copy vector, as a plasmid backbone and used KRX as a host cell. Needless to say, we characterized R0011 in the same experimental condition.For more detailed explanation of the characterization of R0011, go Goal A |
Revision as of 19:15, 25 October 2010
Contents |
Goal B: Characterization of λ Lysis cassette
Introduction
λ Lysis cassette is a gene that causes cell lysis.To characterize the lytic activity of λ Lysis cassette quantitatively, we regulate the gene expression of λ Lysis cassette by a lactose promoter, R0011, which we characterized quantitatively by RPU in GoalA. To repress the basal expression of λ Lysis cassette without IPTG as possible, we used pSB4K5, a low copy vector, as a plasmid backbone and used KRX as a host cell. Needless to say, we characterized R0011 in the same experimental condition.For more detailed explanation of the characterization of R0011, go Goal A
We grew these cultures to saturation at 37 degrees Celsius in supplemented M9 media, and then dillute with the same media 100 fold, growing to mid-log , and split 3ml into falcon tubes. A range of concentrations of IPTG was added to these cultures, and they were then incubated at 37 degrees . and the absorbance at 550nm was measured every 30 min ｗｉｔｈ きかい
