| 摘要: |
| 为深入研究bcl-2对细胞凋亡的调控机制和肿瘤发生发展的影响，应用基因重组手段，根据表达载体pEGFP-Cl上的多克隆位点和bcl-2基因序列设计了1对引物，对含有bcl-2基因的质粒pWB-bcl进行了PCR扩增，得到了708bp的目的片断。将其克隆到pMD18-T vector，筛选阳性克隆并测序，序列分析表明，其与原初序列同源性达99%。将bcl-2插入到载体启动子下游，与报告基因绿色荧光蛋白(Enhancer Green Fluorescent Protein，EGFP)融合，经限制性内切酶酶切和PCR鉴定，证明Bcl-2/EGFP真核表达质粒构建成功。 |
| 关键词: bcl-2 绿色荧光蛋白 真核表达质粒 |
| DOI： |
| 分类号: |
| 基金项目:国家“863”高科技项目（2001AA213081） |
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| Construction of bcl-2/EGFP fused gene eukaryotic expressing plasmid |
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| Abstract: |
| The pro-oncogene B cell lymphoma/leukemia-2 gene (bcl-2) plays an important role in the regulation of apoptosis.In order to study deeply the mechanism,the eukaryotic expressing vector of bcl-2/EGFP fused gene was constructed by the methods of gene recombinating.A pair of primer was designed according to multiple cloning sites of pEGFP-C1 vector and the gene sequence of bcl-2 cDNA.The pWB-bcl plasmid which contains bcl-2 gene was amplified by PCR.The product of PCR was 708 bp.bcl-2 was integrated into pMD18-T vector.The recombinant pMD-bcl plasmid was sequenced and analyzed,homology was 99.bcl-2 gene was inserted into the promoter downstream of pEGFP-C1 vector and fused with Enhancer Green Fluorescent Protein (EGFP) gene.The result affirmed that bcl-2/EGFP fused gene eukaryotic expressing vector was constructed successfully by the restricted endonuclease and PCR. |
| Key words: bcl-2 EGFP fused gene eukaryotic expressing plasmid |
