PubMed:17077011 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PubMed/sourceid/17077011","sourcedb":"PubMed","sourceid":"17077011","source_url":"http://www.ncbi.nlm.nih.gov/pubmed/17077011","text":"[High level expression of recombinant human sCD40L-IZ and identification of its biological activation].\nAIM: To clone and express the recombinant human soluble CD40 ligand(CD40L) with biological activation.\nMETHODS: The isoleucine zipper (IZ) gene was fused at N-terminal of CD40L gene coding E107-L261 contributing to form trimer. The gene coding hexahistidine was fused at the N-terminal of IZ because the sCD40L-IZ fusion protein could be purified by affinity chromatography. Then the fusion gene was amplified and cloned into expression plasmid pET30a.\nRESULTS: The protein sCD40L-IZ with His-tag at the N-terminal was effectively expressed in E. coli BL21 (DE3) as inclusion bodies and a denaturation and refolding procedure was performed to acquire soluble sCD40L-IZ. The fusion protein sCD40L-IZ was conveniently purified using HiTrap affinity column with above 95% purity. The gel filtration chromatography and non-reduced SDS-PAGE identified the trimeric structure of the recombinant protein. The microscope analysis showed the sCD40L-IZ interacted with the membrane CD40 on XG2, a multiple myeloma cell line.\nCONCLUSION: The recombinant human trimeric CD40L-IZ was expressed in prokaryotic cells successfully, which has provided a basis for further study of the relationship between CD40L and apoptosis, CD40L and pathogenesis of illness. It is also useful to clinical treatment.","tracks":[{"project":"Ab3P-abbreviations","denotations":[{"id":"SF0","span":{"begin":172,"end":177},"obj":"ABBR"},{"id":"LF0","span":{"begin":160,"end":171},"obj":"ABBR"},{"id":"SF1","span":{"begin":239,"end":241},"obj":"ABBR"},{"id":"LF1","span":{"begin":220,"end":237},"obj":"ABBR"}],"relations":[{"id":"R0","pred":"ShortForm","subj":"LF0","obj":"SF0"},{"id":"R1","pred":"ShortForm","subj":"LF1","obj":"SF1"}],"attributes":[{"subj":"SF0","pred":"source","obj":"Ab3P-abbreviations"},{"subj":"LF0","pred":"source","obj":"Ab3P-abbreviations"},{"subj":"SF1","pred":"source","obj":"Ab3P-abbreviations"},{"subj":"LF1","pred":"source","obj":"Ab3P-abbreviations"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"Ab3P-abbreviations","color":"#dbec93","default":true}]}]}}