The Construction Of Recombinant Adeno-associated Virus Expressing Human Very Low-density Lipoprotein Receptor Gene

Objective The homo-VLDLR and the recombinant adeno-associated virus expressing VLDLR gene was constructed. Methods Firstly, primers containing specific enzyme-cutting sites was designed and used to amplify a part of the homo-VLDLR from human skeleton muscle and the other part of the homo-VLDLR from a plasmid carrying a part of homo-VLDLR; Secondly, this two part of homo-VLDLR were connected into the complete homo-VLDLR, and the complete homo-VLDLR was cloned into the rAAV-specific plasmid UF1. With the help of plasmid phelper and Pxx2, 293 cell was used to be packing cell, recombinant adeno-associated virus carrying the homo-VLDLR was packed and mensurated its titer。Results human very low-density lipoprotein receptor gene and PXXUF1-VLDLR was successfully constructed, and the titer of rAAV-VLDLR and rAAV-GFP were all 4×1011pfu/ml。Conclusion The titer had arrived to the requestion,so the recombinant adeno-associated virus carrying the homo-VLDLR could be used in gene therapy for hyperlipoidemia in type 2 diabetes...