关键词: 普兰林肽, 密码子突变, 原核表达, 诱导表达

Abstract: Objective To construct the recombination human pramlintide prokaryotic expression vector and induce the expression of pramlintide in E. coli . Methods The human nature pramlintide gene whose genetic codon mutated to prokaryotic preference codon were chemically synthesized. The gene was inserted into pET-39b+ expression vector with double enzyme（ KpnI、HindIII ）digest to construct the recombination plasmid pET-39b+[Pramlintide]，and then，the recombination plasmid was transferred into E. coli BL21 to construct high expression genetic engineering bacteria of pramlintide which was inducted with isopropyl β-D-thiogalactoside（IPTG）. Results Pramlintide was inserted in plasmid at correct site，and gene sequencing indicated that the sequence of pramlintide was the same as exspected. Upon induction with 0. 1 mmol/L IPTG for 3 hours，the recombination protein was expressed in E. coli BL21 cells at 37 ℃ ，The expression of pramlintide reached the highest level and large amounts of recombination protein accumulated in cytoplasm. Conclusion The expression vector pET- 39b + [Pramlintide] was successfully constructed. Upon induction with IPTG，large amounts of recombination protein accumulated in E. coli BL21 cells. It will establish a foundation for producing large amounts of pramlintide in the future.

Key words: pramlintide, codon mutation, prokaryotic expression system, induced expression