Difference between revisions of "Part:BBa K3257044"

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<partinfo>BBa_K3257044 short</partinfo>
 
<partinfo>BBa_K3257044 short</partinfo>
  
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.
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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 a pair of the same lox sites present.
  
 
[[File:Cre+lox Sites.png|none|333px|thumb|'''Fig. 1 The verification of incompatibility between LoxP and different Lox sites.'''
 
[[File:Cre+lox Sites.png|none|333px|thumb|'''Fig. 1 The verification of incompatibility between LoxP and different Lox sites.'''

Revision as of 12:15, 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 a pair of the same 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]