科技创新与应用2024，Vol.14Issue(7)：47-50,4.DOI:10.19981/j.CN23-1581/G3.2024.07.011
四联鲎抗菌肽Tachyplesin Ⅱ毕赤酵母重组质粒构建及其表达
摘要
Abstract
The recombinant Pichia pastoris was used to express quadruple Limulus antimicrobial peptide,Tachyplesin Ⅱ,to achieve efficient preparation.According to the amino acid sequence of Tachyplesin Ⅱ and the specific cleavage sites of pancreatic elastase and carboxypeptidase A,the quadruple(4TpⅡ)sequence was designed and optimized according to the codon preference of Pichia pastoris.After digested by EcoR Ⅰ and Not Ⅰ,the recombinant plasmid pPICZαA-4TpⅡ was constructed and electroporated to Pichia pastoris X-33.The positive transformants were screened by Zeocin resistance and the expression was analyzed by SDS-PAGE.The recombinant plasmid pPICZa A-4Tp Ⅱ was successfully constructed,and the positive transformants were screened.After induced expression at 30℃,250 r/min and 1%methanol for 5 days,the recombinant protein was obtained.This study can lay a foundation for the efficient preparation of Limulus antimicrobial peptide Tachyplesin Ⅱ.关键词
鲎/抗菌肽/Tachyplesin Ⅱ/重组毕赤酵母/串联表达Key words
Limulus/antimicrobial peptide/Tachyplesin Ⅱ/Recombinant Pichia pastoris/tandem expression分类
生物科学引用本文复制引用
曾臻,罗家英,蔡依婕,上官雪茹,黄艺虹,李雅婷,李婉褀,谭强来..四联鲎抗菌肽Tachyplesin Ⅱ毕赤酵母重组质粒构建及其表达[J].科技创新与应用,2024,14(7):47-50,4.基金项目
福建省自然科学基金青创项目(2022J05322) （2022J05322）
福建省中青年教师教育科研项目(JAT210492) （JAT210492）
厦门医学院大学生创新创业训练计划项目(S202112631012,202212631005,202212631006) （S202112631012,202212631005,202212631006）
