GFP reporter gene under the direction of chicken ovalbumin gene promoter expressed in the CHO cell and in the primary cell cultures of chicken oviduct.
- Author:
Yue PANG
1
;
Qing-Wei LI
Author Information
1. College of Life Sciences, Liaoning Normal University, Dalian 116029, China.
- Publication Type:Journal Article
- MeSH:
Animals;
CHO Cells;
Cells, Cultured;
Chickens;
Cricetinae;
Cricetulus;
Epithelial Cells;
cytology;
Female;
Genes, Reporter;
Green Fluorescent Proteins;
biosynthesis;
genetics;
Ovalbumin;
genetics;
Oviducts;
cytology;
Promoter Regions, Genetic;
genetics;
Recombinant Proteins;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2005;21(1):154-158
- CountryChina
- Language:Chinese
-
Abstract:
To reseach GFP reporter gene under the control of chick ovalbumin gene regulatory elements express in the CHO cell and in the primary cell cultures of chicken oviduct. 1.5kb fragment and 2.9kb fragment were amplicated by PCR method, two fragments were subeloned to manmalian expression vector pGFP-N2 by recombinant DNA technology, the CMV promoter was cut off from pGFP-N2, so two expression vectors were constructed, one is the P2.9koval-GFP including promoter, first exon, first intron of chicken ovalbumin gene, the other is the P1.5koval-GFP including first intron of chicken ovalbumin gene. Restriction enzyme digestion and DNA sequence analysis revealed that 5'upstream regions of ovalbumin gene were not only identical to those of the published chicken ovalbumin gene, but also were contained in the recombinant vector. They were transfected into the CHO cell and the primary cell cultures of chicken oviduct by Lipofectin, they were used for fluorescence detection. GFP protein existed in GFP transfected the CHO cell and the primary cell cultures of chicken oviduct. It is demonstrated that GFP reporter gene under the direction of chick ovalbumin gene promoter could be expressed in the CHO cell and in the primary cell cultures of chicken oviduct.
