| pubmed-article:19441235 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:19441235 | lifeskim:mentions | umls-concept:C0086418 | lld:lifeskim |
| pubmed-article:19441235 | lifeskim:mentions | umls-concept:C0007634 | lld:lifeskim |
| pubmed-article:19441235 | lifeskim:mentions | umls-concept:C0017262 | lld:lifeskim |
| pubmed-article:19441235 | lifeskim:mentions | umls-concept:C0162768 | lld:lifeskim |
| pubmed-article:19441235 | lifeskim:mentions | umls-concept:C2248769 | lld:lifeskim |
| pubmed-article:19441235 | lifeskim:mentions | umls-concept:C0054889 | lld:lifeskim |
| pubmed-article:19441235 | lifeskim:mentions | umls-concept:C2911684 | lld:lifeskim |
| pubmed-article:19441235 | lifeskim:mentions | umls-concept:C0185117 | lld:lifeskim |
| pubmed-article:19441235 | pubmed:issue | 1 | lld:pubmed |
| pubmed-article:19441235 | pubmed:dateCreated | 2009-5-15 | lld:pubmed |
| pubmed-article:19441235 | pubmed:abstractText | We constructed the eukaryotic expression vector of human IL-35-IgG4 (Fc)-pOptiVEC-TOPO by gene recombination technique and expressed the fusion protein human IL-35-IgG4 (Fc) in CHO/DG44 cells. The two components of the newly discovered cytokine human IL-35, EBI3 and IL-12p35, were amplified by PCR from the cDNA library derived from the KG-I cells after LPS induction. The two PCR-amplified cDNA fragments of human IL-35 were linked by over-lapping PCR and then cloned into the IgG4 (Fc)-pOptiVEC-TOPO vector. The constructed plasmid with the recombinant cDNA IL-35-IgG4 (Fc) was verified by restriction enzyme digestion analysis, PCR and DNA sequencing. The verified plasmid with the recombinant cDNA was transfected into CHO/DG44 cells using Lipofectamine 2000. The success of the transfection was examined and confirmed by RT-PCR. After selection in alpha-MEM (-) medium, the IL-35-Ig G4 (Fc) positive CHO/DG44 clones were chosen and the media from these positive clones were collected to be used to purify the fusion protein. The positive CHO/DG44 clones were further cultured in increasing concentrations of MTX and the expression levels of the fusion protein IL-35-Ig G4 (Fc) were repetitively induced by MTX-induced gene amplification. The IL-35-IgG4 (Fc) fusion protein was purified from the media collected from the positive CHO/DG44 clones by protein G affinity chromatography and then identified by SDS-PAGE and Western blotting. The results showed that one protein band was found to match well with the predicted relative molecular mass of human IL-35-IgG4 (Fc) and this protein could specifically bind to anti-human IgG4 (Fc) monoclonal antibody. In conclusion, our study successfully established an IL-35-IgG4 (Fc) positive DG44 cell line which could stably express IL-35-IgG4 (Fc) fusion protein. | lld:pubmed |
| pubmed-article:19441235 | pubmed:language | chi | lld:pubmed |
| pubmed-article:19441235 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19441235 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:19441235 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19441235 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19441235 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19441235 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19441235 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:19441235 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:19441235 | pubmed:month | Jan | lld:pubmed |
| pubmed-article:19441235 | pubmed:issn | 1000-3061 | lld:pubmed |
| pubmed-article:19441235 | pubmed:author | pubmed-author:XiaZ FZF | lld:pubmed |
| pubmed-article:19441235 | pubmed:author | pubmed-author:GaoWendaW | lld:pubmed |
| pubmed-article:19441235 | pubmed:author | pubmed-author:ZhangQingQ | lld:pubmed |
| pubmed-article:19441235 | pubmed:author | pubmed-author:ChenYangY | lld:pubmed |
| pubmed-article:19441235 | pubmed:author | pubmed-author:ZhangDaweiD | lld:pubmed |
| pubmed-article:19441235 | pubmed:author | pubmed-author:TangJingJ | lld:pubmed |
| pubmed-article:19441235 | pubmed:author | pubmed-author:LiuQuanshengQ | lld:pubmed |
| pubmed-article:19441235 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:19441235 | pubmed:volume | 25 | lld:pubmed |
| pubmed-article:19441235 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:19441235 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:19441235 | pubmed:pagination | 109-15 | lld:pubmed |
| pubmed-article:19441235 | pubmed:meshHeading | pubmed-meshheading:19441235... | lld:pubmed |
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| pubmed-article:19441235 | pubmed:meshHeading | pubmed-meshheading:19441235... | lld:pubmed |
| pubmed-article:19441235 | pubmed:year | 2009 | lld:pubmed |
| pubmed-article:19441235 | pubmed:articleTitle | [Expression of human IL-35-IgG4 (Fc) fusion protein in CHO/DG44 cells]. | lld:pubmed |
| pubmed-article:19441235 | pubmed:affiliation | College of Life Science, Sichuan University, Chengdu 610041, China. | lld:pubmed |
| pubmed-article:19441235 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:19441235 | pubmed:publicationType | English Abstract | lld:pubmed |
| pubmed-article:19441235 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
