Clone、expression of Human HMGB1 gene and biological activity assays of HMGB1 recombinat protein

China Biotechnology

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Clone、expression of Human HMGB1 gene and biological activity assays of HMGB1 recombinat protein

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Abstract HMGB-1 (high mobility group box－1 protein) gene was amplified by RT-PCR from the mRNA isolated from peripheral blood mononuclear cells (PMBCs) in a patient with liver failure. A recombinant expression vector of GST-HMGB1 was constructed in pGEX4T-1,and transformed into E.coli. It was sequenced and confirmed to be identical to the HMGB-1 gene in data bank(NM_002128). Recombinant protein GST-HMGB1 was purified by Glutathione Sepharose 4B affinity chromatography, and HMGB1 were obtained by thrombin digestion. GST-HMGB1 and HMGB1 were binded to HMGB1's antibody in Western-blotting. From ELISA and MTT assays, the results showed that two proteins may stimulate RAW264.7 cells to secret TNF-α, and induc Hela cells proliferation markedly. GST-HMGB1 had good biological activity and it is employed in further study.

Key words： prokaryotic expression purify immunine activity biological activity Cloning HMGB1

Received: 26 April 2006 Published: 25 November 2006

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. Clone、expression of Human HMGB1 gene and biological activity assays of HMGB1 recombinat protein. China Biotechnology, 2006, 26(11): 20-23.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I11/20

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