重组鸡白细胞介素-7在HEK293T细胞中的表达及其条件分析

Expression of recombinant chicken interleukin-7 in HEK293 Tcells and the condition analysis

为提高重组鸡白细胞介素-7(chIL-7)在真核细胞中的表达水平,本试验分析了影响基因表达的一些因素,包括转染方法、质粒用量、转染时间及表达时间等,对重组chIL-7表达水平的影响,并通过正交试验设计方法确定了利用HEK293T细胞表达重组chIL-7的最适表达条件。结果表明,在T75细胞瓶中,由脂质体介导使用30μg质粒,转染4h,表达48h和由磷酸钙介导使用40μg质粒,转染6h,表达60h可以获得较高的表达效率,表达水平分别为(124±9)和(94.3±8)μg/106个细胞。本结果为进一步研究chIL-7的功能提供有利条件。

To enhance expression level of recombinant chicken interleukin-7（chIL-7）in eukaryotic cells,here we analyzed the effects of some factors involved in gene expresssion in eukaryotic cells on chIL-7expression.The factors included transfection methods,vector doses,transfection time and expression time.On this basis,we used the orthogonal test to determine the optimum expression condition in HEK293 Tcells.Results showed that the high-level expression was achieved in T75 flask using 30μg vectors for 4htransfection and 48 hexpression mediated by liposome,and using 40μg vectors for 6htransfection and 60 hexpression mediated by calcium phosphate,respectively.The expression levels were（124 ± 9）μg/106 cells and（94.3 ± 8）μg/106 cells in liposome and calcium phosphate mediations,respectively.Thisstudy provided necessary conditions for further investigation on chIL-7.