人白细胞介素-15的高效表达

Study on the High Expression of Human Interleukin-15

关键词:  大肠杆菌,IL-15基因,基因表达,优化

Key words:  Escherichia coli,IL-15 gene,Gene expression,optimization

摘要:

采用限制性核酸内切酶酶切鉴定含IL-15基因的重组质粒,SDS-PAGE检测不同条件下IL-15基因的表达情况,并对表达条件进行优化。经酶切鉴定和核苷酸序列测定证实重组质粒pET28c( )-IL-15含有IL-15基因且阅读框架正确,以IPTG诱导IL-15基因表达的优化条件是：培养基pH 7.0,培养温度37℃,IPTG浓度1.0mmol/L,菌体生长密度OD600达到1.0时加入IPTG,诱导时间3h,此时IL-15蛋白表达量为28%。从而实现了IL-15基因的高效表达并优化了其表达工艺。从而为IL-15的生物活性和生产工艺研究提供了可靠的试验数据。

Abstract:

The recombinant plasmid pET28c( )-IL-15 was identified with restriction endonucleases digestion and the expressional level of differently conditional IL-15 protein was detected by SDS-PAGE. The expression conditions were optimized. By identification of endonucleases digestion and sequence analysis, the recombinant expression plasmid pET28c( )-IL-15 contained IL-15 gene having positive reading frame. The expression optimization result indicated that the IL-15 gene expression optimized condition with IPTG induction is culture medium pH 7.0,culture temperature 37℃, joining IPTG to final concentration 1.0mmol/L when the recombinant strain growth density OD600 achieved 1.0, and induction time 3h. The expression level of the IL-15 protein were about 28% of total cellular protein with IPTG induction by SDS-PAGE and gel system analysis. The IL-15 protein is expressed high by recombinant strain BL21(DE3)( pET28c( )-IL-15). The expression technology was optimized. This provided experimental data to research the biology activity and productive technology of IL-15.