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OALib Journal期刊
ISSN: 2333-9721
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Couple Production of Human Calcitonin and Rat Peptidylglycine αamidation Monooxygenase in Insect Cells
人修饰型降钙素和鼠酰胺化酶基因在昆虫细胞中的偶联表达

Keywords: 降钙素,酰胺化酶,偶联表达,杆状病毒

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Abstract:

Human calcitonin (hCT) is a 32 amino acid peptide hormone that requires C-terminal amidation for full biological activity. Calcitonin has important physiological function in vivo. We describe the couple expression of a synthesized modified human calcitonin(hmCT) gene fused with glutathione-S-transferase and rat peptidylglycine alpha-amidation monooxygenase (PAM) in insect cells infected by recombinant baculovirus GSTCT/PAM. Using Western blotting against hmCT or rat PAM, the GSThmCT fusion protein had been identified as well as the PAM. Following affinity chromatography with glutathione agarose column, the GSThmCT fusion protein produced by insect cells was purified. The purified fusion protein was also interacted with antibody against hmCT. The couple expression of a modification enzyme and its substrate in eucaryotic expression system may be used for producing other biological activity peptides.

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