Aquaporin 4-siRNA interfering sequence, its fusion expression vectors and medicinal use of the fusion expression vectors
An aquaporin and interfering sequence technology, applied in the field of aquaporin 4-siRNA interfering sequence and its fusion expression vector, can solve the problems that are not yet fully clarified, and achieve the effects of inhibiting invasion and infiltration and inhibiting tumor spread
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[0026] 1. Sources of experimental materials:
[0027] Human malignant glioma cell line LN-229 was provided by the University of Texas MD Anderson Cancer Center, the chemotaxis chamber was purchased from Neuro Probe, EGF was purchased from Peprotech, the intersectin1 antibody was from Abcam, and G418 was purchased from Invitrogen company.
[0028] 2. Design and synthesis of AQP4-siRNA
[0029] Design specific intersectinl-siRNA according to the siRNA design software provided by Ambion on the Internet, and synthesize the following templates by Shanghai Gemma Pharmaceutical Technology Co., Ltd., with BamH I and Bbs I restriction sites at both ends, and siRNA action sites: 5'-GGCTGGCTTGGAGGAGAATTA-3'.
[0030] 3. Preparation of AQP4-siRNA fusion expression vector
[0031] The expression vector pGPU6 / GFP / Neo vector has a linear structure with sticky ends of BamH I and Bbs I restriction sites at both ends. The synthesized AQP4-siRNA was annealed, connected into the pGPU6 / GFP / Neo...
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