Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/32558
Title: Expression of carcinoscorpius rotundicauda factor C in eukaryotes
Authors: DING, JEAK LING 
HO, BOW 
Issue Date: 12-Jan-1999
Source: DING, JEAK LING,HO, BOW (1999-01-12). Expression of carcinoscorpius rotundicauda factor C in eukaryotes. ScholarBank@NUS Repository.
Abstract: CrFC21 cDNA was cloned into two mammalian vectors: pCIneo and pCDNAI, both of which carry the strong CMV promoter for expression in mammalian cell lines. Various CrFC cDNA constructs transformed into P. pastoris and S. cerevisiae were expressed to yield full-length recombinant Factor C (rCrFC) protein of .about.130 kDa which is immunoreactive. The rCrFC is expressed in an intracellular, insoluble form. Intracellular localization of the nascent protein provides protection from premature digestion by proteases secreted by the host cell. Subsequent to its synthesis, rCrFC is solubilized and purified under pyrogen-free conditions. Using established protocols, the protein can be denatured and renatured to recover its biological functionality. By manipulation of the 5' end of CrFC26, truncated constructs containing this cDNA are expressed by S. cerevisiae to give immunoreactive rCrFC. The rCrFC produced from both CrFC21 and CrFC26 constructs, solubilized by Triton X-100 or SDS, is found to be immunoreactive. Solubilized rCrFC was purified as a proenzyme and reversibly protected from activation by addition of Me.sub.2 SO.
URI: http://scholarbank.nus.edu.sg/handle/10635/32558
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
US5858706.PDF731.52 kBAdobe PDF

OPEN

PublishedView/Download

Page view(s)

147
checked on Dec 8, 2017

Download(s)

63
checked on Dec 8, 2017

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.