作　　者： ; ; ; ; ; ; ; ; (叶琬）;

机构地区： 成都医学院

出　　处： 《徐州医学院学报》 2008年第3期141-144,共4页

摘　　要： 目的克隆人白细胞介素27(interleukin 27,IL-27)蛋白亚基EB病毒诱导的基因3(Epstein-Barr vi-rus-induced gene 3,EBI3)与p28基因的cDNA,分别构建其真核表达质粒并在COS-7细胞表达。方法人外周血单核细胞诱导为成熟树突状细胞后提取细胞总RNA,经逆转录-聚合酶链反应(RT-PCR)扩增EBI3与p28基因cDNA,酶切后插入pcDNA-3.1构建真核表达载体pcDNA-EBI3与pcDNA-p28。重组质粒转染COS-7细胞,RT-PCR检测目的基因表达。结果成功构建克隆人IL-27蛋白亚基真核表达质粒,重组质粒分别转染COS-7细胞后检测到相应基因表达。结论人IL-27蛋白亚基基因克隆及真核表达质粒的成功构建,为构建表达有生物学活性的重组人单链IL-27蛋白奠定基础。 Objective To clone and construct the eukaryotic expression plasmids containing human IL- 27 protein subunits and investigate their expression following transfection into COS -7 cells. Methods Total RNA was extracted from dendritic cells derived from human peripheral blood monocytes for cloning EBI3 cDNA and p28 cDNA by RT - PCR. The eukaryotic expression plasmids pcDNA - EBI3 and pcDNA - p28 were constructed and transfected into COS - 7 cells, where the expressions of target gene were detected by RT - PCR. Results The IL - 27 protein subunits cDNA were cloned and the constructed plasmids were verified to be correct by sequencing. Target gene expressions were detected by RT -PCR in the transfected COS -7 cells. Conclusion The eukaryotic expression plasmids containing human IL -27 protein subunits were constructed and expressed in COS -7 cells successfully. This work paves the way for construction and expression of the bioactive single chain human IL - 27 protein.

关 键 词： 基因克隆 真核表达

领　　域： [生物学]