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haofurui新虫 (正式写手)
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[求助]
英译汉，谢谢
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Recombinant human ANX protein was expressed using a commercial ANX clone (Genecopeia, Germantown, MD). Competent BL21 E. Coli were transformed with a DNA plasmid containing a glutathione-S-transferase (GST)- ANX (human) fusion cDNA and a 50-T7 bacterial promoter; colonies were cultured overnight in 100 mM Ampicillin/10 mM IPTG medium; and plasmids were purified (Qiaprep Miniprep, Alameda, CA), confirmed by PCR (forward-50-GCTGGCAAGCCACGTTTGG-30, reverse- 50-TTCACTTCTGAGTTCGGCATG–30), grown overnight(250 mL), and isolated (Maxiprep, Qiagen). The ANXGST fusion protein was purified on glutathione-agarose affinity columns (Thermo Scientific, Waltham, MA), confirmed by SDS-PAGE (Bio-Rad, Hercules, CA), and cleaved with enterokinase to remove the GST tag. Free ANX was purified by passing through a glutathione-agarose affinity column a second time to remove free GST and quantified by Bradford protein assay. |
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