[Reconstruction and expression of green fluorescent protein and aquaporin 7 fusion recombinant vector].
OBJECTIVE
To 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.


METHODS
The 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.


CONCLUSIONS
The CHO cell line with stable pEGFP-C1-AQP7 expression was set up successfully for advanced research.
Yang
Ke
K
Department of Reproductive Pathophysiology, School of Medical Sciences, Jilin University, Changchun, Jilin 130021, China.
Li
Feng
F
Zhao
Dan
D
Li
Danan
D
Liu
Qingshuang
Q
Liu
Xichun
X
Zhang
Jian
J
Zhao
Xuejian
X
Yang
Baoxue
B
D000818:Animals; D020346:Aquaporins / Q000096:biosynthesis* / Q000235:genetics; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D018076:DNA, Complementary / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013737:Testis / Q000378:metabolism*; D014162:Transfection
C114141:AQP7 protein, human; D020346:Aquaporins; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins