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《Acta Universitis Medicinalis Nahui》 2002-01
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Construction of pBK- human non-muscle MLCK N-terminal and its expression in E. coli XL1-blue

Hu Ruolei, Zhu Huaqing, Zhou Qing et al (Laboratory of Molecular Biology, Anhui Medical University, Hefei 230032)  
Objective To construct the recombinant vector for expression of human non-muscle myosin light chain kinase (hnmMLCK) N-terminal in E.coli. Methods PCR was used to amplify hnmMLCK N-terminal cDNA, which was inserted into pBK-cmv to construct expression vector. The recombinant vector was transformed into XL1-blue and IPTG was used to induce expression. SDS-PAGE and electroelude were used to purify expressed product. Results Restriction endonucleases mapping analysis and DNA sequencing demonstrated that the constructed plasmid was the recombinant one. The expressed protein was purified by electroelude, and the quantity is about 21% of the whole protein. Conclusion The expressed vector is successfully constructed and hnmMLCK expressed in E.coli, which lay a good basis for manufacture and clinical application of the enzyme.
【Fund】: 国家自然科学基金 (编号 :39870 32 4);; 教育部优秀年轻教师基金;; 安徽省自然科学基金 (编号 :990 44312 );; 安徽省教育厅自然科学基金 (编号 :99JL0 0 90 )
【CateGory Index】: R346
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