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Bulletin of Botanical Research ›› 2011, Vol. 31 ›› Issue (1): 56-60.doi: 10.7525/j.issn.1673-5102.2011.01.010

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Expression,Purification and Immunoreactivity of Hygromycin-B-Phosphotransferase

GAO Bin;ZHANG Hai-Yan;FAN Hai*   

  1. 1.Key Laboratory of Plant Stress,Shangdong Normal University,Jinan　250014；2.Institute of Botany,the Chinese Academy of Sciences,Beijing　100093
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-01-20 Published:2011-01-20
  • Contact: FAN Hai
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Abstract: The prokaryotic expression plasmid of hygromycin-B-phosphotransferase(HPT) was constructed. The hpt gene was cloned by PCR. It was digested by SalⅠ/NotⅠand subcloned into expression vector pET-28b(+). pET-28b-hpt was transferred into E. coli Rosset; fusion protein was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) and analyzed by SDS-PAGE. The most products existed in an inclusion body form. The HPT protein purified by Ni2+-NTA column was used to immunize New Zealand rabbits. The HPT polyclonal antibodies reveal high sensitivity and specificity.

Key words: hygromycin-B-phosphotransferas, prokaryotic expression, recombinant protein, polyclonal antibodies

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