Reconstruction and expression of green fluorescent protein and aquaporin 7 fusion recombinant vector / 中华男科学杂志

ABSTRACT

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>(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.</p><p><b>CONCLUSION</b>The CHO cell line with stable pEGFP-C1-AQP7 expression was set up successfully for advanced research.</p>