Difference between revisions of "Part:BBa K4585008"
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Revision as of 11:35, 9 October 2023
pGL4.35-3xHA-9xGAL4UAS-KRAB-NLS linearized vector
The objective was to synthesize the pGL4.35-3 HA-9 GAL4UAS-KRAB-NLS plasmid by homologous recombination with the KRAB homologous recombination fragment .
pGL4.35-3×HA-9×GAL4UAS-KRAB-NLS linearized vector
We based on the sequence of the pGL4.35 plasmid. Through homologous recombination experiments, the Gal 4 sequence, 3× HA sequence were ligated to the pGL4.35 plasmid, and nine UAS sequences were inserted simultaneously. Special primers were designed, and then the plasmid was cut into linear sequences of a length of about 4500 bp by PCR. The two ends of the linear sequence can be complementary to both ends of the KRAB homologous recombination fragment, and the purpose is to perform homologous recombination with the KRAB homologous recombination fragment to obtain the target product pGL4.35-3×HA-9×GAL4UAS-KRAB-NLS plasmid.
1 Pattern Diagram
Fig.1 The agarose gel electrophoresis of pGL4.35-3×HA-9×GAL4UAS-KRAB-NLS linearized vector
2 Experiment
2.1 Method
Based on the sequence of the pGL4.35-3 plasmid, we designed the pGL4.35-3×HA-9×GAL4UAS-KRAB-NLS linearized vectror by using the GAL 4 homologous recombination fragment, 9×UAS homologous recombination fragment and homologous recombination experiments.
2.2 Results
Fig.1 The model diagram of pGL4.35-3×HA-9×GAL4UAS-KRAB-NLS
3.Caution
The two ends of the pGL4.35-3×HA-9×GAL4UAS-KRAB-NLS linearized vectror need to be complementary to the two ends of the corresponding recombinant fragments to ensure the success of homologous recombination ligation.