摘要
目的构建Der p2真核表达载体,并证明能在小鼠骨髓来源的树突状细胞(DC)中表达。方法采用分子生物学方法,将原核表达质粒plambd Der p2携带的Der p2全长cDNA切下,重组到真核表达质粒pCI-neo中,然后在脂质体介导下,将重组质粒转染到小鼠骨髓来源的DC,并用RT-PCR、Western Blot检测Der p2 mRNA和蛋白表达。结果测序证实构建的重组质粒中携带了Der p2的全长cDNA序列,且与Gene Bank序列完全一致;重组质粒转染小鼠DC后,能产生Der p2 mRNA和蛋白。结论成功构建重组Der p2基因真核表达载体,其转染DC后,能有效地表达于DC中。
Objective To construct Der p2 gene eukaryotic expression vector, and to detect the expression after transfecting into dendritic cells（DC）. Methods Complete Der p2 cDNA was spliced from prokaryotic expression vector plambd-Der p2, and then cloned into eukaryotic expression vector pCbneo （pCI-neo-Der p2）. The positive recombinants pCI-neo-Der p2 transfected into DC, then RT-PCR was used to detect the expression of Der p2 mRNA, and Western Blot assayed the expression of Der p2 protein. Results Sequencing result showed Der p2 cDNA in pCI-neo-Der p2 was in coincidence with the sequence registrated in Gene Bank. In the DC after transfection of pCI-neo-Der p2, the result of RT-PCR showed that Der p2 mRNA expressed, and Western Blot also detected the expression of Der p2 protein. Conclusion The Der p2 cDNA was constructed into the eukaryotic expression vector successfully, and Der p2 gene and protein could be expressed efficiently in DC.
出处
《重庆医学》
CAS
CSCD
2006年第11期997-999,共3页
Chongqing medicine
基金
国家自然科学基金资助项目(30470772)
