To enhance porcine interleukin-7 (pIL-7) expression in HEK293T cells, pIL-7 gene was codon-optimized according to the human preferred codons. The eukaryotic expression vector of the pIL-7 gene fused with Myc/His-tag was generated using pcDNA3.1A plasmid. For pIL-7 expression, the codon-optimized pIL-7 vector and wild-type pIL-7 vector constructed previously were transfected into HEK293T cell, respectively, the recombinant pIL-7 was purified from the culture media with Ni-agrose beads, and the pIL-7 expression levels from the two vectors was compared. Results showed that the expression level of codon-optimized pIL-7 gene was 45 μg/T25 flask, more than twice as much as its wild-type gene (21 μg/T25 flask), indicating that codon optimization significantly enhances pIL-7 expression in HEK293T cells. This study provided a necessary condition for further investigation on biological functions of porcine IL-7.%为提高重组猪白细胞介素-7(pIL-7)在人胚胎肾细胞(HEK293T)中的表达,依据人偏爱的密码子对pIL-7基因进行了优化修饰,然后用pcDNA3.1A质粒构建其与Myc/His-标签融合的真核表达载体.将此表达载体和过去构建的野生型pIL-7表达载体分别转染HEK293T细胞进行表达,利用镍-琼脂糖凝胶颗粒从培养基中纯化表达的重组pIL-7,比较二者的表达水平.结果显示,密码子优化后pIL-7基因在HEK293T细胞中的表达水平为(45±3.2) μg/T25细胞瓶,比野生型pIL-7基因的表达水平(21±1.8) μg/T25细胞瓶,提高了2倍多,可见密码子优化可明显提高重组pIL-7在HEK293T细胞中的表达.这为猪IL-7的生物学功能的研究提供了必要条件.
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