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dc.contributor.authorLuo, Zhen
dc.contributor.authorXu, Zhinan
dc.contributor.authorZhuo, Shanling
dc.contributor.authorJing, Keju
dc.contributor.authorLu, Yinghua
dc.contributor.author卢英华
dc.date.accessioned2013-04-19T08:33:34Z
dc.date.available2013-04-19T08:33:34Z
dc.date.issued2012-03-15
dc.identifier.citationBIOCHEMICAL ENGINEERING JOURNAL,2012,62:86-91zh_CN
dc.identifier.issn1369-703X
dc.identifier.urihttp://dx.doi.org/10.1016/j.bej.2012.01.002
dc.identifier.uriWOS:000301966000012
dc.identifier.urihttps://dspace.xmu.edu.cn/handle/2288/16149
dc.description.abstractHuman Fas ligancl (hFasL) is a type II membrane protein that induces apoptosis in the Fas-bearing cells. Its special biological activity has the potential for the therapeutic use as an anti-cancer agent directed at enhancing apoptosis in tumor cells. In this study Escherichia coli and eukaryotic Dictyostelium discoideum were used to produce a soluble form of hFasL in large amounts. An expression vector for hFasL production in E. coli was constructed based on plasmid pET32a(+). By cultivation of the hFasL-producing E. coli clone on LB medium and induction with IPTG, a hFasL concentration of 1.0 mg L-1 was achieved. D. discoideum strain AX3-hFasL-H was cultured in a conventional stirred bioreactor on an improved synthetic medium using a simple fed-batch strategy, and cell densities of up to 8.3 x 10(7) cells/mL and a maximum hFasL concentration of 420 mu g/L were obtained. Using Ni-NTA affinity chromatography purification, two kinds of recombinant hFasLs from E. coli and D. discoideum were purified with a purity of 94% and 90%, respectively. They showed similar biological activities in inducing apoptosis in Fas-expressing cells. (C) 2012 Elsevier B.V. All rights reserved.zh_CN
dc.description.sponsorshipProgram for New Century Excellent Talents in Fujian Province, PR China; National Natural Science Foundation of China [31071488, 20736008, 20928006]zh_CN
dc.language.isoenzh_CN
dc.publisherELSEVIER SCIENCE SAzh_CN
dc.subjectHuman Fas ligandzh_CN
dc.subjectEscherichia colizh_CN
dc.subjectDictyostelium discoideumzh_CN
dc.subjectSoluble productionzh_CN
dc.subjectAffinity chromatographyzh_CN
dc.subjectCytotoxic activityzh_CN
dc.titleProduction, purification and cytotoxity of soluble human Fas ligand expressed by Escherichia coli and Dictyostelium discoideumzh_CN
dc.typeArticlezh_CN


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