Reconstruction and expression of green fluorescent protein and aquaporin 7 fusion recombinant vector.
- Author:
Ke YANG
1
;
Feng LI
;
Dan ZHAO
;
Danan LI
;
Qingshuang LIU
;
Xichun LIU
;
Jian ZHANG
;
Xuejian ZHAO
;
Baoxue YANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Aquaporins; biosynthesis; genetics; CHO Cells; Cricetinae; Cricetulus; DNA, Complementary; genetics; Green Fluorescent Proteins; biosynthesis; genetics; Male; Molecular Sequence Data; Rats; Rats, Wistar; Recombinant Fusion Proteins; biosynthesis; Reverse Transcriptase Polymerase Chain Reaction; Testis; metabolism; Transfection
- From: National Journal of Andrology 2004;10(11):819-823
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the membrane mobility of aquaporin 7 (AQP7) by cloning stably transfected CHO cells with expression of pEGFP-C1-AQP7, in which AQP7 cDNA was fused downstream and in frame to pEGFP-C1 gene.
METHODSThe full sequence of AQP7 was amplified by RT-PCR and then recombined in the downstream of the green fluorescent protein gene in the pEGFP-C1 vector. The recombinant vector pEGFP-C1-AQP7 was stably transfected into CHO cells. With fluorescent microscopy, immunocytochemical stain and Western blot, pEGFP-C1-AQP7 showed a predominant intracellular vesicular localization.
RESULTS(1) The sequence of AQP7 cDNA of the Wistar rat was logged into the GenBank (access number: AY157737). (2) Identification demonstrated that pEGFP-C1-AQP7 fusion protein stably expressed in CHO cells. (3) With fluorescence microscopy, pEGFP-C1-AQP7 showed a predominant intracellular vesicular localization.
CONCLUSIONThe CHO cell line with stable pEGFP-C1-AQP7 expression was set up successfully for advanced research.
