Details

Autor(en) / Beteiligte

• Liu, Yi-Qing
• Chen, Zi-Jiang
• Zhang, Xue
• Wang, Lai-Cheng
• Jiao, Yu-Lian
• Zhang, Jie
• Ma, Chun-Yan
• Cui, Bin
• Gao, Xin-Pu
• Liu, Zheng-Min
• Wu, Kan
• Zhao, Yue-Ran

Titel

Cloning and expression of human interleukin-26 in Escherichia coli

Ist Teil von

• Shengwu gongcheng xuebao, 2006-05, Vol.22 (3), p.413

Ort / Verlag

China

Erscheinungsjahr

2006

Quelle

MEDLINE

Beschreibungen/Notizen

• To clone human interleukin-26 (hIL-26) and express it in E. coli efficiently. Two pairs of primers were synthesized according to the hIL-26 gene reported on GenBank. The hIL-26 gene was cloned by nest PCR following the first round RT-PCR from human peripherial blood monocytes total RNA, and then the PCR product was cloned into pMD18-T vector. Colony PCR, restriction analysis and sequence analysis showed that the gene cloned was the same as the reported hIL-26. The recombinant was cut with BamHI and EcoR I to obtain the hIL-26 fragment, and then the fragment was inserted into pBV220 which was cut with the same enzymes. The recombinant expression vector was induced to express hIL-26 at 42 degrees C, SDS-PAGE analysis showed that the recombinant protein accounted for up to 20% of the whole protein of E. coli, and the protein was also confirmed by Western blotting. Purity of the protein was found to be above 90% after purified with molecular sieve. After renaturalized with glutathione buffer, the promoting effect