Team:PKU Beijing/Project/AND Gate 2 Design
From 2009.igem.org
(Difference between revisions)
(→One of the Second AND Gate Design) |
(→One of the Second AND Gate Design) |
||
Line 14: | Line 14: | ||
- | In order to creat a amber mutation inside of the PhiR73 delta/T3pol coding sequence, we carried out site directed mutagenesis with primers introduced with TAG amber mutation at the original Ser site. One problem that comes to us is how to decide which serine codon to mutate. In order to avoid that the half translated protein has function, we picked first few serine of the whole sequence of either PhiR73 delta activator and T3 polymerase. For PhiR73 delta it is on the first and second serine. (For PhiR73 delta sequence click [http://partsregistry.org/Part:BBa_I746352 Here]) | + | In order to creat a amber mutation inside of the PhiR73 delta/T3pol coding sequence, we carried out site directed mutagenesis with primers introduced with TAG amber mutation at the original Ser site. One problem that comes to us is how to decide which serine codon to mutate. In order to avoid that the half translated protein has function, we picked first few serine of the whole sequence of either PhiR73 delta activator and T3 polymerase. For PhiR73 delta it is on the first and second serine. (For PhiR73 delta sequence click [http://partsregistry.org/Part:BBa_I746352 Here]). In T3 polymerase, we picked four sites, they are on the 37th, 91th, 130th, 283th nucleotide of its coding sequence. The Sequence of the T3 polymerase with an amber mutation on its 37th and 91th nucleotide can be found in the parts that we hand in this year. |
+ | |||
[[Image:PKU_Point_Mutation.png|500px|center|thumb|fi2. Creating the Amber mutation]] | [[Image:PKU_Point_Mutation.png|500px|center|thumb|fi2. Creating the Amber mutation]] | ||
Revision as of 17:17, 13 October 2009