猪瘟病毒流行毒株E2基因密码子优化及在酵母中的高效表达

Codon Optimization and Expression in Pichia pastoris of E2 Gene of Classical Swine Fever Virus

密码子的偏嗜性是影响基因异源表达的重要参数之一 ,优化密码子序列能够提高外源基因的表达水平。野生型猪瘟病毒E2基因在毕赤酵母中的表达量较低 ,为了提高E2基因在毕赤酵母中的表达水平 ,利用PCR基础上的定点突变技术将E2基因上的毕赤酵母低利用密码子突变为其高利用率的简并密码子。结果表明 ,替换野生型E2基因上 2 4个酵母低利用密码子后 ,E2基因在酵母中表达水平有较显著提高 。

Codon bias was one of the important parameter which influence heterogenous gene expre- ssion, optimizing codon sequence could improve expression level of he teroge nous gene. In the preview study, wildtype E2 gene was expressed poorly in Pich ia pastoris, in order to improve the expression level of E2 gene in Pichia pas tori s, the low usage codons of E2 gene were mutated into high usage codons in Pi chia pastoris by directed-mutagenesis based on PCR. The result showed that,compa re d with the results reported in preview study, the expression level of E2 gene in Pichia pastoris was improved observably by substituting 24 low usage codons of E2 gene for the high usage synonymous codons. It suggested the stragety to impro ve the expression of E2 gene in Pichia pastoris by codon optimization was su ccessful.