Construction and expression of recombined human AFP eukaryotic expression vector

来源 :World Journal of Gastroenterology | 被引量 : 0次 | 上传用户:beanmilk
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AIM:To construct a recombined human AFP eukaryoticexpression vector for the purpose of gene therapy and targettherapy of hepatocellular carcinoma (HCC).METHODS:The full length AFP-cDNA of prokaryotic vectorwas digested,and subcloned to the multi-clony sites of theeukaryotic vector.The constructed vector was confirmed byenzymes digestion and electrophoresis,and the productexpressed was detected by electrochemiluminescence andimmunofluorescence methods.RESULTS:The full length AFP-cDNA successfully cloned tothe eukaryotic vector through electrophoresis,0.9723 IU/ml AFP antigen was detected in the supernatant of AFP-CHO by electrochemiluminescence method.Compared withthe control groups,the differences were significant (P<0.05).AFP antigen molecule was observed in the plasma of AFP-CHO by immunofluorescence staining.CONCLUSION:The recombined human AFP eukaryoticexpression vector can express in CHO cell line.It providesexperimental data for gene therapy and target therapy ofhepatocellular carcinoma. AIM: To construct a recombined human AFP eukaryoticexpression vector for the purpose of gene therapy and target of hepatocellular carcinoma (HCC). METHODS: The full length AFP-cDNA of prokaryotic vectorwas digested, and subcloned to the multi-clony sites of theeukaryotic vector. The constructed vector was confirmed byenzymes digestion and electrophoresis, and the productexpressed was detected by electrochemiluminescence and immunofluorescence methods .RESULTS: The full length AFP-cDNA successfully cloned tothe eukaryotic vector through electrophoresis, 0.9723 IU / ml AFP antigen was detected in the supernatant of AFP- CHO by electrochemiluminescence method. Compared with the control groups, the differences were significant (P <0.05). AFP antigen molecule was observed in the plasma of AFP-CHO by immunofluorescence staining. CONCLUSION: The recombined human AFP eukaryoticexpression vector can express in CHO cell line .It providesexperimental data for gene therapy and target therapy of hepatocellular ca rcinoma.
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