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中文摘要：
为了构建由双CMV启动子调控的真核双顺反子表达载体pcDNA3.1AB,首先在pcDNA3.1（＋）的基础上,构建出pcDNA3.1A和pcDNA3.1B,然后通过PCR方法扩增pcDNA3.1A上含PCMV-MCS-BGHpolyA表达单元的片段,再定向克隆入pcDNA3.1B,最后得到载体pcDNA3.1AB。将DsRed与EGFP基因插入pcDNA3.1AB,分别构建pcDNA-DsRed、pcD-NA-EGFP和pcDNA-DsRed-EGFP真核表达质粒,并转染COS-7细胞瞬时表达。用荧光显微镜进行检测,结果表明,双价质粒pcDNA-DsRed-EGFP能同时表达红色和绿色荧光蛋白基因,两基因表达强度无显著差异,且双价与单价质粒相比荧光蛋白表达强度相当。
英文摘要：
To construct a bieistronic expression vector pcDNA3.1AB controlled by two CMV promoters, pcDNA3.1A and pcDNA3.1B were constructed first by reforming the MCS of pcDNA3.1（＋）. The PCR product of PCMV-MCS-BGHpolyA from pcDNA3. 1A was inserted into pcDNA3.1B to form pcDNA3.1AB. DsRed gene and EGFP gene were inserted into pcDNA3.1AB to construct pcDNA-DsRed, pcDNA-EGFP and pcDNA-DsRed-EGFP. The recombinants were transfected and transiently expressed in COS-7 cells and detected by fluorescence microscope. DsRed and EGFP were coexpressed in pcDNA-EGFP-DsRed, and the fluorescence intensity had no significant deviation. The fluorescence intensity was similar between monocistronic and bicistronic vectors.
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