| 杨继鑫 王洋杨 魏洪亮 陈聪 杨玉庆 李信 李南林.14-3-3θ基因过表达载体的构建及其过表达乳腺癌细胞株的建立[J].,2015,15(15):2820-2823 |
| 14-3-3θ基因过表达载体的构建及其过表达乳腺癌细胞株的建立 |
| Vector Construction of Over Expressed Gene 14-3-3θand Its Breast CancerCell Line Build |
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| DOI： |
| 中文关键词: 14-3-3θ pcDNA4.0 基因重组技术 乳腺癌 |
| 英文关键词: 14-3-3θ pcDNA 4.0 Recombinant DNA technology Breast cancer |
| 基金项目:国家自然科学基金项目（810721179） |
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| 中文摘要: |
| 目的：为进一步研究14-3-3θ基因对乳腺癌生物学行为的影响，构建14-3-3θ基因真核细胞过表达重组载体并建立其过表达
的乳腺癌细胞株。方法：合成的14-3-3θ基因重组在pENTR 3C DUAL载体中，课题组采用DNA重组技术将14-3-3θ基因剪切分
离后，插入真核表达质粒载体pcDNA4.0 中，重组得到真核过表达载体pcDNA4.0-14-3-3θ，并用其转染293T 细胞后用western
blot进行验证，将经过验证的真核过表达载体pcDNA4.0-14-3-3θ转染乳腺癌细胞后用zeocin 筛选，筛选出的乳腺癌细胞经过
western-blot和PCR 验证确为稳定过表达14-3-3θ的乳腺癌细胞。结果：成功构建14-3-3θ基因过表达真核载体并建立其过表达乳
腺癌细胞株，揭示14-3-3θ基因可能在乳腺癌的早期诊断和预后预测中具有重要作用。 |
| 英文摘要: |
| Objective:In order to further study the effect of biology behavior of the gene 14-3-3θto breast cancer, to build a recombinant
vector and breast cancer cell line which can over express gene 14-3-3θ.Methods:The gene 14-3-3θin the pENTR 3C DUAL is
synthesized by a company, we put the gene 14-3-3θinto the vector pcDNA4.0 which can product 14-3-3θin eukaryotic cells by recombinant
DNA technology and then transfect the 293T, we identify the overexpression of the recombinant vector by the western blot and
PCR. At last, we transfect the breast cancer cell by the recombinant vector and screen the breast cancer cell with Zeocin.Results:Conclusion:Successfully
construct the recombinant vector and breast cancer cell line which can over express 14-3-3θ. Which shows that gene 14-3-3θ
may play an important role in the early diagnosis and prognosis of breast cancer prediction. |
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