Difference between revisions of "Part:BBa K3257044"
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| − | Cre recombinase is responsible for recombination of reverse transcribed cDNA, which is also one of the most important processes in our mutation library construction. We expressed Cre recombinase in E.coli and | + | 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. |
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| + | [[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.]] | ||
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| − | <span class='h3bb'>Sequence and Features</span> | + | <span class='h3bb'>'''Sequence and Features'''</span> |
<partinfo>BBa_K3257044 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3257044 SequenceAndFeatures</partinfo> | ||
Latest revision as of 12:08, 13 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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 356
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 42
- 1000COMPATIBLE WITH RFC[1000]