人CC10基因在肺腺癌A549细胞中的表达
Expression of Human CC10 Gene in Lung Adenocarcinoma A549 Cell
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摘要: 目的构建及鉴定人CC10基因真核细胞表达载体。方法采用RT-PCR法,从人肺组织的总RNA中扩增出273bp的人CC10cDNA片段,利用DNA重组技术将其克隆到真核细胞表达载体pcD-NA3.1中,脂质体法转染pcDNA3.1-hCC10到肺腺癌A549细胞,荧光免疫细胞化学法及Westernblot法检测CC10蛋白的表达。结果用酶切重组质粒并行序列鉴定,人CC10基因已经正确克隆到真核细胞表达载体pcDNA3.1中,体外染pcDNA3.1-hCC10的肺腺癌A549细胞中CC10蛋白表达明显增高。结论成功构建了CC10基因的真核细胞表达载体pcDNA3.1-hCC10,并获得表达Abstract: Objective To construct and identity human the expression plasmid pcDNA3.1-hCC10. Methods A 273 bp cDNA fragment was amplified from the total RNA of normal lung tissue by RT-PCR,then inserted it into expression vector pcDNA3.1 by DNA recombinant techniques . We transfected pcDNA3.1- hCC10 in A549 cells by liposome-mediated gene transfer method. The protein expression of CC10 was detected by the techniques of immunofluorescence and Western blot. Results After the identification of restriction enzymes analysis...
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