Application of FBXW7 or up-regulator thereof in preparing medicament for treating diabetes and preventing and treating tumorigenesis of diabetic individuals
A technology of diabetes and diabetic blood vessels, applied in the field of biomedicine, can solve the problem of not equal to diabetes and obesity mouse models.
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[0081] 1. Method
[0082] 1. Construction of viral vector overexpressing EWS (name: E-OE) or Fbxw7 (name: F-OE) protein
[0083] (1) Amplify the nucleotide sequences of EWS and Fbxw7 by PCR reaction (the EWS sequence is shown in SEQ ID NO: 1, and the Fbxw7 sequence is shown in SEQ ID NO: 2), and the two ends are connected to EcoRI and BamHI restriction sites point;
[0084] (2) Double-enzyme digestion PCR product and purification;
[0085] (3) H201pAdeno-MCMV-EGFP-3FLAG vector plasmid (Shanghai Heyuan Biotechnology Co., Ltd.) was also double-enzyme digested, the large fragment was recovered and purified, and the EWS or Fbxw7 gene fragment in step (2) was inserted by molecular cloning technology instead of EGFP to the EcoRI and BamHI restriction sites of the shuttle plasmid;
[0086] (4) Screening transformants by colony PCR, and the screened positive clones are verified by sequencing;
[0087] (5) Sequencing to verify the correct clone, and perform high-purity plasmid extr...
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