west china medical publishers
Author
  • Title
  • Author
  • Keyword
  • Abstract
Advance search
Advance search

Search

find Author "LIUXiaoyun" 1 results
  • Construction of cTnC-linker-TnI(P) Genes, Expression of Fusion Protein and Preparation of Lyophilized Protein

    In order to construct and express human cardiac troponin C-linker-troponin I(P) fusion protein, detect its activity and prepare lyophilized protein, we searched the CDs of human cTnC and cTnI from GenBank, synthesized cTnC and cTnI(30-110aa) into cloning vector by a short DNA sequence coding for 15 neutral amino acid residues. pColdⅠ-cTnC-linker-TnI(P) was constructed and transformed into E. coli BL21(DE3). Then, cTnC-linker-TnI(P) fusion protein was induced by isopropyl-β-D-thiogalactopyranoside (IPTG). Soluable expression of cTnC-linker-TnI(P) in prokaryotic system was successfully obtained. The fusion protein was purified by Ni2+ Sepharose 6 Fast Flow affinity chromatography with over 95% purity and prepared into lyophilized protein. The activity of purified cTnC-linker-TnI(P) and its lyophilized protein were detected by Wondfo FinecareTM cTnI Test. Lyophilized protein of cTnC-linker-TnI(P) was stable for 10 or more days at 37℃and 4 or more months at 25℃and 4℃. The expression system established in this research is feasible and efficient. Lyophilized protein is stable enough to be provided as biological raw materials for further research.

    Release date: Export PDF Favorites Scan
1 pages Previous 1 Next

Format

Content