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中文摘要：
目的：提高肌钙蛋白Ⅰ（troponinⅠ，TnⅠ）在大肠杆菌中的可溶性表达并进行分离纯化。方法和结果：将质粒CBD—intein／pTWIN1和TnⅠ／pET28a分别用NcoⅠ及XhoⅠ双酶切，回收。回收后的载体CBD—intein／pTWIN1与TnⅠ片段用T4连接酶连接，得到重组质粒CBD—intein—TnⅠ／pTWIN1。将重组质粒转化大肠杆菌BL21（DE3），经IPTG诱导表达得到融合蛋白CBD—intein-TnⅠ。经chitin亲和层析和intein介导的融合蛋白自我裂解，纯化得到重组TnⅠ。结论：通过CBD—intein融合表达系统一步式裂解分离纯化能方便快捷地获得可溶性的TnⅠ。
英文摘要：
Objective To highly express soluble recombinant troponin Ⅰ （Tn Ⅰ ） in E. coli and obtain purified protein-Tn Ⅰ. Method and Result The Tn Ⅰ gene was obtained by digesting the plasmid Tn Ⅰ -pET28a using restriction enzymes Nco Ⅰ and Xho Ⅰ ,the purified Tn Ⅰ was inserted into Nco Ⅰ / Xho Ⅰ sites of CBD-intein/pTWⅠN1 to form a recombinant expression plasmid CBD-intein-Tn Ⅰ/pTWⅠN1, then transformed it into E. coli for expression. The fusion protein CBD-intein-Tn Ⅰ was expressed by addition of TPTG and purified by chitin affinity chromatography. Conclusion The soluble Tn Ⅰ can conveniently obtain by CBD-intein expression system and chitin chromatography.
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