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| 人白细胞介素-15的高效表达 | |
| 引用本文： | 许崇利.人白细胞介素-15的高效表达[J].中国兽药杂志,2012,46(4):7-9. |
| 作者姓名： | 许崇利 |
| 作者单位： | 1. 吉林化工学院环境与生物工程学院,吉林吉林132022／吉林大学畜牧兽医学院,长春130062 2. 吉林化工学院环境与生物工程学院,吉林吉林,132022 3. 大连大学医学院,辽宁大连,116622 |
| 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pH7．0，培养温度37℃，IPTG浓度1．0mmol／L，茵体生长密度0D600达到1．0时加入IPTG，诱导时间3h，IL-15蛋白表达量为28％，为IL-15的生产工艺研究提供了可靠的试验数据。 |
| 关 键 词： | 大肠杆菌 IL-15基因 基因表达 优化 |
| 收稿时间： | 1/2/2012 6:03:10 PM |
| 修稿时间： | 2012/2/10 0:00:00 |
Study on the High Expression of Human Interleukin-15 |
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| XU Chong-Li.Study on the High Expression of Human Interleukin-15[J].Chinese Journal of Veterinary Drug,2012,46(4):7-9. | |
| Authors: | XU Chong-Li |
| Institution: | College of Environment and Bioengineering,Jilin Institute of Chemical Technology,Jilin |
| 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 expression plasmid pET28c（ ＋ ） - IL - 15 contained IL - 15 gene having positive reading frame. The result indicated that the IL - 15 gene expression optimized condition is culture medium pH 7.0, culture temperature 37 ℃, joining IPTG to final concentration 1.0 mmol,/L when the recombinant strain growth density OD600 achieved 1.0, and induction time 3 h. The expression level was 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 productive technology of IL - 15. |
| Keywords: | Escherichia coli IL-15 gene Gene expression optimization |
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