Targetted liver cancer AFP gene siRNAs expression carrier and its constructing method and use
A technology for expressing vectors and genes, used in the fields of siRNAs expression plasmids and gene therapy drugs
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[0071] The present invention will be further described below in conjunction with embodiment, but not limited thereto.
[0072] 1. Materials used for the preparation of siRNAs expression vector targeting liver cancer AFP gene
[0073] 1.1 Plasmids and strains Plasmid pSilencer 3.0-H1 was purchased from Ambion, see figure 1 , the engineering strain DH5α was purchased from the Institute of Molecular Biology of Shandong University.
[0074] 1.2 Enzymes and main reagents T4 DNA ligase was purchased from Promega Company in the United States. The extraction and purification of plasmids were prepared by classic alkaline lysis method and polyethylene glycol plasmid purification method.
[0075] 1.3 Sequencing primers M13 universal primers were used for sequencing the inserts in the plasmids. Forward Primer: 5'-CACGACGTTGTAAAAGAC-3', Reverse Primer: 3'-GGACACATTTAACAATAG G-5', purchased from Shanghai Shenyou Company.
[0076] 1.4 Cell line and medium: The liver cancer cell line SMMC-...
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