%0 Journal Article
%T Expression and Purification of Recombinant Human Interleukin 4 in Escherichia coli<br>重组人IL-4大肠杆菌表达与纯化
%A QIU Yan
%A SUN Jiu-Ru
%A HUANG Yang-Bin
%A HUANG Zhi-Hua
%A CHEN Lin-Jie
%A LIU Yong
%A LIN Yue-Xin
%A <br>邱燕
%A 孙九如
%A 黄阳滨
%A 黄智华
%A 陈霖杰
%A 刘勇
%A 林跃鑫
%J 生物工程学报
%D 2006
%I 
%X Human interleukin 4 (IL-4) cDNA was optimized and synthesized according to E. coli preferred codon. A recombinant expression plasmid pET-30a (+)/rhIL-4 was constructed with the target cDNA inserted between Nde I and EcoR I sites, which can translate the mature IL-4 protein with an extra methionine residue at N-terminal. The expression vector was transformed into E. coli BL21 (DE3). The rhIL-4 protein was expressed in the inclusion body. By using the optimized fermentation conditions, the high expression level was achieved with the expression level as high as 35% of total protein obtained. A purification strategy has been designed which includes Q-Sepharose and SP-Sepharose ion-exchange chromatography and dialysis renaturation. The rhIL-4 was purified with the purity more than 98% and the yield of 40 mg per liter fermentation culture achieved. Western blot proved that the purified protein is IL-4. Amino acid sequencing revealed that N-terminal 16 residue sequence is identical to the theoretical sequence. Biological activity assay on TF-1 cells demonstrated that the rhIL-4 is active with an activity of 2.5 x 10(6) AU/mg. This study promises large scale production of rhIL-4.
%K rhIL-4
%K Escherichia coli
%K fermentation
%K purification<br>rhIL-4
%K 大肠杆菌
%K 发酵
%K 纯化
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=3927BC21F3E1C956&yid=37904DC365DD7266&vid=BC12EA701C895178&iid=B31275AF3241DB2D&sid=CFC2B32D03D9F610&eid=F131C0ADC94A25CF&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=19