Codon Preference Optimization Increases Prokaryotic Cystatin C Expression.
Gene expression is closely related to optimal vector-host system pairing in many prokaryotes. Redesign of the human cystatin C (cysC) gene using the preferred codons of the prokaryotic system may significantly increase cysC expression in Escherichia coli (E. coli). Specifically, cysC expression may...
| Publicado en: | Journal of Biomedicine & Biotechnology Vol. 2012; pp. 1 - 8 |
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| Autores principales: | , , , |
| Formato: | pictorial research tables/charts Journal Article |
| Publicado: |
Wiley-Blackwell
2012
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=104297850&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 104297850 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 11107243 137K jtl: Journal of Biomedicine & Biotechnology issn: 11107243 maglogo: N pubinfo: dt: 2012 vid: 2012 pid: 480 pub: Wiley-Blackwell place: Malden, Massachusetts artinfo: ui: 104297850 104297850 2011906772 NLM23093857 PMC3471025 104297850 ppf: 1 ppct: 7 formats: fmt: @attributes: type: P tig: atl: Codon Preference Optimization Increases Prokaryotic Cystatin C Expression. aug: au: Qing Wang Cui Mei Honghua Zhen Jess Zhu affil: Department of Clinical Laboratory, Affiliated Hospital of Qingdao University Medical College, Qingdao 266003, China sug: subj: Genetic Engineering Methods Recombinant Proteins Analysis Gene Expression Escherichia Coli Recombinant Proteins Metabolism Human Sequence Analysis Reverse Transcriptase Polymerase Chain Reaction Blotting, Western Funding Source ab: Gene expression is closely related to optimal vector-host system pairing in many prokaryotes. Redesign of the human cystatin C (cysC) gene using the preferred codons of the prokaryotic system may significantly increase cysC expression in Escherichia coli (E. coli). Specifically, cysC expression may be increased by removing unstable sequences and optimizing GC content. According to E. coli expression system codon preferences, the gene sequence was optimized while the amino acid sequence was maintained. The codon-optimized cysC (co-cysC) and wild-type cysC (wt-cysC) were expressed by cloning the genes into a pET-30a plasmid, thus transforming the recombinant plasmid into E. coli BL21. Before and after the optimization process, the prokaryotic expression vector and host bacteria were examined for protein expression and biological activation of CysC. The recombinant proteins in the lysate of the transformed bacteria were purified using Ni2+-NTA resin. Recombinant protein expression increased from 10% to 46% based on total protein expression after codon optimization. Recombinant CysC purity was above 95%. The significant increase in cysC expression in E. coli expression produced by codon optimization techniques may be applicable to commercial production systems. pubtype: Academic Journal doctype: pictorial research tables/charts Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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