上海农业学报2024，Vol.40Issue(4)：81-85,5.DOI:10.15955/j.issn1000-3924.2024.04.13
密码子优化和信号肽对猪圆环病毒ORF2基因在毕赤酵母表达中的影响
Impact of codon optimization and signal peptide on the expression of PCV2 ORF2 gene in Pichia pastoris
摘要
Abstract
In order to efficiently express porcine circovirus 2(PCV2)ORF2 gene in Pichia pastoris,the codons of 187 amino acid of PCV2 ORF2 novel phytase gene was chemically synthesized by the PCR-based two-step DNA synthesis according to yeast codon preference.Unoptimized or optimized PCV2 ORF2 genes,with or without encoding signal peptide were inserted into pPICZα-C expression vector.The recombinant plasmids were linearized and transformed into Pichia pastoris GS115.Recombinant strains were preliminarily selected by Zeocin resistance.Only the PCV2 ORF2 gene,which was optimized gene and removed encoding signal peptide,was successfully expressed.The expression of recombinant protein was approved by SDS-PAGE and Western-blot analysis.The molecular weight of the target protein is about 30 ku.That the recombinant expressed protein could be recognized by anti-PCV2 serum which means the recombinant ORF2 protein had bioactivity.关键词
猪圆环病毒ORF2基因/密码子优化/稀有密码子/自身信号肽/毕赤酵母Key words
Porcine circovirus 2(PCV2)ORF2 gene/Codon optimization/Rare codon/Native signal peptide/Pichia pastoris分类
农业科技引用本文复制引用
张春玲,王瑞阳,钱忠辉,赵本进,张俊平,葛宇燕,张子悦,丁卫星..密码子优化和信号肽对猪圆环病毒ORF2基因在毕赤酵母表达中的影响[J].上海农业学报,2024,40(4):81-85,5.基金项目
上海市科技兴农项目(2022-02-08-00-12-F01454) （2022-02-08-00-12-F01454）
