毕传林 刘海艳 韩翠芳 张信玲 顾玉超.稳定表达 p120ctn 的人肺腺癌 A549 细胞株的构建[J].现代生物医学进展英文版,2015,15(2):213-216. |
稳定表达 p120ctn 的人肺腺癌 A549 细胞株的构建 |
The Construction of Human Lung Adenocarcinoma Cell Line of A549Expressing p120ctn |
|
DOI: |
中文关键词: p120ctn 肺癌 A549 细胞株 稳定高表达 |
英文关键词: p120ctn Lung adenocarcinoma A549 cell line Overexpression |
基金项目:国家自 然科学基金项目( 81 172013); 高等学校博士学科点专项科研基金资助课题( 201 001 32120012) |
|
Hits: 1118 |
Download times: 1066 |
中文摘要: |
目 的: 构建稳定表达 p120ctn 的 A549 细胞株,以研究 p1 20ctn 蛋白在肺癌发生和转移过程中的作用。方法: 通过分子克隆, 将
pcDNA3.1 多 克隆位点插入 Flag 标签的编码序列 ,得到 pcDNA.Flag 表达载体。 然后 PCR 扩增 p1 20ctn 的编码序列 , 插入 Flag 标
签下游,构建 pcDNA.Flag-p1 20ctn 质粒, 筛 选阳性克隆并进行酶切及测序鉴定。 利 用 脂质体 Lipofectamine 2000 将 pcDNA.
Flag-p1 20ctn 质粒转染到 肺癌细胞 A549 中,通过 G418 筛选得到 稳定转染细胞株, 免疫印迹法检测 p1 20ctn 的表达。 结果: 本文构
建了融合有 Flag 标签的 p120ctn 真核表达载体并转染到 A549 中, 免疫印迹结果表明 p120ctn 蛋白在 A549 细胞中高效的表达。
结论: 本文成功构建了稳定高表达 p1 20ctn 的 A549 细胞模型, 为深入研究 p1 20ctn 在肺癌的发生和转移过程中的作用 奠定了基
础。 |
英文摘要: |
Objective:To establish a stable A549 cell line over-expressing p120ctn through pcDNA3.1.Methods:First, a DNA
fragment encoding Flag-tag was synthesized. A p1 20ctn gene was cloned by PCR and inserted into the multiple cloning sites of pcDNA3.
1 to construct the plasmid pcDNA.Flag-p120ctn. The vector was identified by enzyme digestion and DNA sequencing. Secondly, we
transfected pcDNA.Flag-p120ctn into A549 cell by lipofectamine 2000. Then, stable transfected A549 cell line expressing p120ctn was
established after screening culture by G41 8 and was identified by Western blot.Results:The plasmid pcDNA. Flag-p120ctn was
constructed. Stable transfected A549 cell line was established and p1 20ctn was over-expressed successfully.Conclusion:We successfully
generate a stable A549 cell line expressing p1 20ctn which sets a cell model for further research into the genesis and development of
human lung adenocarcinoma. |
View Full Text
View/Add Comment Download reader |
Close |