鸡肌球蛋白轻链激酶在大肠杆菌中的表达

Expression of chicken myosin light chain kinase in escherichia coli

目的建立鸡肌球蛋白轻链激酶（ｍｙｏｓｉｎｌｉｇｈｔｃｈａｉｎｋｉｎａｓｅ，ＭＬＣＫ）ｃＤＮＡ表达克隆。方法将ＭＬＣＫ功能区（自动抑制区、钙调蛋白识别区和活性催化区）的ｃＤＮＡ插入质粒ｐＢＫｒｓｖ中，重组质粒（ｐＢＫｒｓｖ－ＭＬＣＫ）转化入大肠杆菌中并获得表达。结果Ｗｅｓｔｅｒｎ印迹试验表明表达产物的分子量与插入片段ｃＤＮＡ所含信息相一致。结论所获重组克隆在体外表达。

Objective The expressive clone of chicken myosin light chain kinase (MLCK) had been constructed. Methods The pBKrsvMLCK had been subcloned the functional sequence (autoinhibition, calmodulin recognization and activity regions) of MLCK into plasmid pBKrsv polylinker, and transformed into E coli XL1blue and expressed. Results Western blot analysis and MLCK activity measurement showed that the product expressed contains all information of MLCK cDNA inserted. Conclusions MLCK has been expressed and has activity of peptide substract phosphorylation.