【摘要】 目的构建黑色素瘤抗原基因-12(MAGE-12)绿色荧光蛋白表达载体pEGFP-C3-MAGE-12,并在真核细胞中表达。方法于2005-10～2005-12对郑州大学第一附属医院应用RT-PCR方法,从人肺癌组织中扩增出MAGE-12cDNA基因片段,经过酶切鉴定后,克隆至质粒载体(pGEM-Teasy),测序证实碱基序列无误后,再克隆至真核绿色荧光蛋白表达载体(pEGFP-C3)上,并转染真核细胞,观察其在真核细胞中表达。结果经酶切及基因序列分析验证,PCR扩增出944bp的MAGE-12基因并成功构建了真核绿色荧光蛋白表达载体pEGFP-C3-MAGE-12,该重组载体能够在真核细胞中广泛表达。结论成功构建真核绿色荧光蛋白表达载体pEGFP-C3-MAGE-12,为建立肿瘤细胞疫苗打下基础。更多还原

【Abstract】 Objective To construct eukaryotic fluorescent expression vector pEGFP-C3 -MAGE-12 and induce the vector express in eukaryotic cells.Methods From Oct. 2005 to Dec. 2005,mRNA was extracted from human lung cancer pecimen,MAGE-12 gene fragment was amplified by reverse transcriptase polymerase chain reaction(RT-PCR). After the amplified product was examined by electrophoresis and sequence determination,this fragment was inserted into pEGFP-C3 fluorescent expression vector .Then the expression vector was transfected into eukaryotic cell. Results The restriction enzyme digestion and sequence analysis showed that recombined cloning vector pGEM-T-MAGE -12 and expression vector pEGFP-C3-MAGE-12 had been constructed successfully,which can express in eukaryotic cell stably.Conclusion Constructing the expression vector pEGFP-C3-MAGE-12 successfully,which make the foundation for producing a new tumor cell vaccine.更多还原