目的　构建人非肌肉肌球蛋白轻链激酶 (hnmML CK)N端表达载体及其体外表达与表达产物的纯化。方法　用PCR方法扩增hnmMLCK的N端cDNA ,插入质粒 pBK cmv中构建成表达载体 ,转化入大肠杆菌XL1 blue ,经IPTG诱导表达 ,表达产物用电洗脱的方法初步纯化。结果　经限制性内切酶酶切图谱分析和DNA序列测定证明所构建质粒为hnmMLCK重组质粒 ,SDS PAGE证实获得分子量为 2 1ku的融合蛋白 ,表达量约占菌体总蛋白的 2 1%。结论　成功构建了重组hnmMLCK表达载体 ,并在大肠杆菌中获得表达 ,为进一步研究及临床应用奠定了良好基础 Objective To construct a N-terminal expression vector of human non-myosin light chain kinase (hnmML CK) and its expression in vitro and purification. Methods The N-terminal cDNA of hnmMLCK was amplified by PCR and inserted into plasmid pBK cmv to construct the expression vector. The recombinant plasmid was transformed into E. coli XL1 blue and induced by IPTG. The expressed product was purified by electroelution. Results Restriction endonuclease digestion and DNA sequencing proved that the constructed plasmid was hnmMLCK recombinant plasmid. The fusion protein with the molecular weight of 21 ku was obtained by SDS PAGE, and the expression level was about 21% of the total bacterial protein. Conclusion The recombinant hnmMLCK expression vector was successfully constructed and expressed in E. coli, which laid a good foundation for further research and clinical application