Expression of Human Stem Cell Factor Mimetic Peptides Fused with c-jun Leucine Zipper in <em>E.coli</em> and Identification of Its Bio-activity

China Biotechnology

2010, Vol. 30

Issue (12): 5-9 DOI:

Expression of Human Stem Cell Factor Mimetic Peptides Fused with c-jun Leucine Zipper in E.coli and Identification of Its Bio-activity

SU Lin¹, KONG Yan², LIU Chang-zheng¹, YANG Ke-gong¹, CHEN Song-sen¹

1. National Laboratory of Medical Molecular Biology,Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, Beijing 100005, China;
2. Department of Renal Cancer and Melanoma, Peking University School of Oncology, Beijing Cancer Hospital & Institute, Beijing 100036, China

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Abstract

The two human stem cell factor (hSCF) mimetic peptides (CS2 and LS2) fused with the c-jun leucine zipper respectively were expressed in E.coli and the bio-activity of the fusion proteins was identified. The three expression plasmids pET30a-CSJ, pET30a-LSJ and pET30a-Jun, which contain fusion proteins (CSJ and LSJ) and c-jun gene respectively, were constructed by using overlapping PCR. Results showed that the CSJ, LSJ and c-Jun(CJ )were expressed about 20% of total protein in E.coli BL21. After purified by using affinity Ni-NTA and Sephadex G-50 column chromatography, the purity of the CSJ, LSJ and CJ were over 95% identified by using SDS-PAGE analysis. Their molecular mass was 7336.0 8, 7991.54 and 6672.74 respectively determined by mass spectrometry. The bioactivity of the fusion proteins was evaluated by using cell proliferation assay with MTT in UT-7 cell. Compared to CS2 and LS2, the bioactivities of CSJ and LSJ were dramatically improved and it is about 1000 fold higher than that of CS2 and LS2.

Key words： Human stem cell factor Mimetic peptide Fusion protein Leucine zipper

Received: 23 July 2010 Published: 25 December 2010

ZTFLH:

R542.220.5

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	SU Lin
	KONG Yan
	LIU Chang-zheng
	YANG Ke-gong
	CHEN Song-sen

Cite this article:

SU Lin, KONG Yan, LIU Chang-zheng, YANG Ke-gong, CHEN Song-sen. Expression of Human Stem Cell Factor Mimetic Peptides Fused with c-jun Leucine Zipper in E.coli and Identification of Its Bio-activity. China Biotechnology, 2010, 30(12): 5-9.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2010/V30/I12/5


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