融合蛋白GST-PADI4可溶性表达条件的优化及纯化  

Optimization of condition for soluble expression of GST-PADI4 fusion protein and purification of expressed product

作　　者：柴政斌[1,2] 张更林[1] 王学政[1,2] 韩金祥[1] 

机构地区：[1]卫生部生物技术药物重点实验室山东省罕少见病重点实验室山东省医药生物技术研究中心山东省医学科学院,山东济南250062 [2]济南大学山东省医学科学院医学与生命科学学院,山东济南250062

出　　处：《中国生物制品学杂志》2014年第3期404-408,411,共6页Chinese Journal of Biologicals

摘　　要：目的优化融合蛋白GST-PADI4的可溶性表达条件,并对蛋白进行纯化。方法对融合蛋白GST-PADI4工程菌的诱导温度(16、20、25、30、37℃)、诱导剂IPTG浓度(0.05、0.1、0.2、0.5、1 mmol/L)、诱导时菌液A600值(0.2、0.4、0.6、0.8、1.0)、诱导时间(8、12、16、20、24 h)进行优化,分析融合蛋白GST-PADI4的可溶性表达情况;按优化的表达条件进行大量表达后,采用Sepharose 4B亲和层析柱对融合蛋白进行纯化。结果在16℃,菌液A600值约为0.4时,以0.1 mmol/L IPTG诱导12 h,融合蛋白GST-PADI4有较高的可溶性表达;纯化的融合蛋白纯度为91%。结论优化了融合蛋白GST-PADI4的可溶性表达条件,并获得了高纯度的可溶性融合蛋白,为后续研究PADI4蛋白的功能奠定了基础。Objective To optimize the condition for soluble expression of GST-PADI4 fusion protein and purify the expressed product. Methods The temperature for induction （16, 20, 25, 30 and 37 °C）, concentration of IPTG as an inducer （0. 05, 0. 1, 0. 2, 0. 5 and 1 mmol/L）, Am value of bacterial liquid at time of induction （0. 2, 0. 4, 0. 6, 0. 8 and 1.0） and time for induction （8, 12, 16, 20 and 24 h） of recombinant E. coli were optimized, and the soluble expression of GST- PADI4 was analyzed. The fusion protein was expressed in a large quantity under the optimal condition and purified by Sepharose 4B affinity chromatography. Results The condition for soluble expression of GST-PADI4 fusion protein was optimized as induction of bacterial liquid at an A600 value of about 0. 4 with 0. 1 mmol/L IPTG at 16 ~C for 12 h. The fusion protein was highly expressed in a soluble form and reached a purity of 91% after purification. Conclusion The condition for soluble expression of GST-PADI4 fusion protein was optimized, and soluble fusion protein with high purity was obtained, which laid a foundation of further study on the function of PADI4 protein.

关 键 词：肽酰基精氨酸脱亚胺酶Ⅳ 融合蛋白 可溶性表达 纯化 

分 类 号：Q786[生物学—分子生物学]