Construction and identification of a eukaryotic expression vector for APP695 gene containing green fluorescence protein gene
10.3760/cma.j.issn.1671-8925.2008.12.002
- VernacularTitle:携带绿色荧光蛋白基因的APP695真核细胞表达载体的构建及鉴定
- Author:
Jing LIANG
1
;
Ning LI
;
Fen-Bin WANG
;
Li-Juan ZHANG
;
Ji-Yu JU
Author Information
1. 潍坊医学院
- Keywords:
Alzheimer's disease;
APP695;
Eukaryotic expression vector
- From:
Chinese Journal of Neuromedicine
2008;7(12):1193-1195
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a eukaryofic expression vector for A PP695 gene carrying green fluorescence protein (GFP) gene. Methods RT-PCR was used to amplify the full-length APP695 cDNA. The PCR products and the eukaryotic expression vector pIRES2-EGFP were digested by restriction endonueleases, and the digested APP695 gene was inserted into the digested eukaryotic expression vector. The positive recombinants were identified by PCR analysis, Nhe Ⅰ and Hind Ⅲ digestion and sequence analysis. Results The 2088-bp DNA fragment was amplified by PCR from the plasmid pCB6, and an identical DNA fragment was also amplified from the recombinants. The products of double restriction enzyme digestion were A PP695 gene with a 5.3-kb DNA fragment. Sequence analysis confirmed successful insertion of A PP695 gene into pIRES2-EGFP vector. Conclusion The eukaryotie expression vector pIRES2/APP695-EGFP has been successfully constructed.
