MBP-1分子真核表达载体的构建及其在食管癌细胞株Eca109中的高表达

目的:初步探讨MBP-1的分子功能.方法:通过PCR扩增获得MBP-1基因编码序列,将其定向插入质粒pcDNA3.1(+)构建重组质粒pcDNA3.1-mbp-1,重组质粒转染体外培养的食管癌Eca109细胞株,Western-blotting检测MBP-1在食管癌细胞中的表达.结果:筛选出分子量较大的重组质粒,酶切分...

Construction of the Eukaryotic Expressing Vector for MBP-1 and Its Expression in Esophageal Cancer Cells

Objective： To preliminarily study the molecular function of MBP-1 molecule.Methods： The MBP-1 gene fragment was obtained by polymerase chain reaction（PCR）.The MBP-1 gene was inserted into plasmid pcDNA3.1（＋） to construct the recombinant eukaryotic expressing vector for MBP-1.Esophageal cancer cells were transfected with a recombinant vector.MBP-1 expression was detected by using Western-blotting.Results： A recombinant plasmid was obtained during screening.The recombinant plasmid contained the coding sequence of MBP-1 gene,which was identified by restriction enzyme analysis and DNA sequencing.Western-blotting analysis showed that MBP-1 was overexpressed in esophageal cancer cells with recombinant plasmid transfection.Conclusions： A recombinant eukaryotic expressing vector for MBP-1 was successfully constructed.MBP-1 was over expressed in esophageal cancer cells after transfection with the recombinant plasmid.These results provide the basis for further study of the molecular function of MBP-1.