Cloning of human APOE ?3 gene and construction of its eukaryotic expression vector
- VernacularTitle:人载脂蛋白E?3等位基因的克隆及表达
- Author:
Li GUI
- Publication Type:Journal Article
- Keywords:
Apolipoprotein E;
Alleles;
Gene clone;
Mammalian expression
- From:
Journal of Chongqing Medical University
2007;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To clone human APOE?3 gene for construction of mammalian expression vector pEGFP-N1-APOE?3. Methods:The cDNA encoding APOE?3 was cloned by the methods RT-PCR from total human embryonic brain RNA. The gene was ligated with PMD19-T vector and then was excised from PMD19-T-APOE?3 plasmid and inserted into pEGFP-N1 to construct eukaryotic expression vector. The expression of APOE?3 gene was identified by western-blot. Results:APOE?3 gene sequence and vector were identified correctly by enzyme digestion and sequence analysis. The APOE?3 gene was transfected into HEK293 cells and expressed successfully. Conclusion:The APOE?3 gene is successfully cloned and its eukaryotic expression vector is obtained,which provides basis for the further experimental researches on APOE functions.
