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mazhaohua

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[求助] 帮忙翻译一下这句话,感谢 已有1人参与

Recombinant OmpA22-350 (rOmpA) protein was expressed with hexahistidine affinity tags at their N termini using the expression vector TAGZyme pQE2 (Qiagen Sciences,
Germantown, MD). The gene fragment was amplifiedusing PCR and Pfu polymerase (Promega KK, Tokyo, Japan), E. coli strain ATCC 25922T genomic DNA as the template, and the primers Fw_rOmpA22 and Rv_rOmpA350. The PCR fragments were cloned into pQE2 at the SalI and XhoI sites, and the resulting plasmids were transformed into E. coli BL21 (DE3) (GE Healthcare, Tokyo, Japan) for protein expression. The recombinant proteins were purified using Ni2 + -chelate chromatography.
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wanglei512

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【答案】应助回帖

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mazhaohua: 金币+6, ★★★很有帮助 2015-05-13 19:33:20
通过在OmpA22-350 (rOmpA)基因N末端融合6个His(组氨酸)标签,运用TAGZyme pQE2表达载体表达了重组蛋白OmpA22-350 (rOmpA)。以大肠杆菌ATCC 25922T基因组DNA为模板,用引物对Fw_rOmpA22 / Rv_rOmpA350和Pfu聚合酶,PCR扩增了基因片段。将PCR扩增得到的基因片段,通过SalI 和 XhoI酶切位点,克隆到pQE2质粒中,然后转化大肠杆菌BL21 (DE3)感受态细胞,进行蛋白表达分析。重组蛋白用镍柱螯合层析法进行纯化。
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