Difference between revisions of "Part:BBa K3257044"
Line 4: Line 4:
 
Cre recombinase is responsible for the recombination of reverse-transcribed cDNA, which is also one of the most important processes in our mutation library construction. We have expressed Cre recombinase in E.coli and proved its ability of gene cleavage when there are lox sites present.
 
Cre recombinase is responsible for the recombination of reverse-transcribed cDNA, which is also one of the most important processes in our mutation library construction. We have expressed Cre recombinase in E.coli and proved its ability of gene cleavage when there are lox sites present.
  
−
[[File:ILacI+IPTG.png|none|500px|thumb|'''Figure 2. The induction level of EGFP under different repressors and promoters.''' The induction level is calculated by dividing the fluorescence level after 9 h of induction by 1 h afterward. The fluorescence level is quantified as in Fig. 1. oLacI stands for the wildtype LacI, iLacI stands for our improved version of LacI. +LacO indicates that the promoter constitutes a LacO sequence. t-test analysis shows that the induction level of iLacI is significantly higher than oLacI, *** indicates that p=0.0002.]]
+
[[File:Cre+lox Sites.png|none|333px|thumb|'''Fig. 1 The verification of incompatibility between LoxP and different Lox sites.'''
 +
Plasmids containing the Cre-SRC gene were co-transformed respectively with plasmids containing mCherry flanked by a LoxP site and either LoxP, Lox2272 or Lox5171 site. A pair of sequencing primers were used to amplify the mCherry gene. PCR product would be around 1000 bp while fragments cleaved by Cre recombinase would be approximately 250 bp. The positive control group is the PCR product of plasmids containing the original mCherry gene. The negative control group is the PCR product of plasmids containing the Cre-SRC gene only.]]
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 12:14, 11 October 2019

Cre Recombinase

Cre recombinase is responsible for the recombination of reverse-transcribed cDNA, which is also one of the most important processes in our mutation library construction. We have expressed Cre recombinase in E.coli and proved its ability of gene cleavage when there are lox sites present.

Fig. 1 The verification of incompatibility between LoxP and different Lox sites. Plasmids containing the Cre-SRC gene were co-transformed respectively with plasmids containing mCherry flanked by a LoxP site and either LoxP, Lox2272 or Lox5171 site. A pair of sequencing primers were used to amplify the mCherry gene. PCR product would be around 1000 bp while fragments cleaved by Cre recombinase would be approximately 250 bp. The positive control group is the PCR product of plasmids containing the original mCherry gene. The negative control group is the PCR product of plasmids containing the Cre-SRC gene only.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 356
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 42
  • 1000
    COMPATIBLE WITH RFC[1000]