摘要
为了利用组成型表达方法在Pichia pastoris SMD1168中表达抗菌肽Cecropin AD,利用化学方法合成编码抗菌肽Cecropin AD的基因片段SH,将基因片段SH插入到载体pGAPZa-A中命名为pGAPZa-ASH,将pGAPZa-ASH电转至Pichia pastoris SMD1168中,Zecion抗性筛选阳性重组子。用Tricine-SDS-PAGE分析表达状况,琼脂扩散法检测蛋白的抑菌活性。结果表明:在3.0ku处有目的条带,并且表达产物抗菌肽Cecropin AD有生物活性。研究利用组成型表达方法在毕赤酵母菌株成功表达了抗菌肽Cecropin AD。组成型表达方法在本研究中成功的应用,为以后的实验研究奠定了理论基础。
To take constitutive expression method that antibacterial peptide Cecropin AD was expressed in Pichia pastoris SMD1168.Gene fragments SH encoding antimicrobial peptides Cecropin AD was synthesized by chemical methods.Gene fragment SH was inserted into the vector pGAPZa-A and named pGAPZa-ASH.Plasmid PGAPZa-ASH was converted to Pichia pastoris SMD1168 using electrotransformation.Screened positive recombinant by Zecion resistance.The expression was analysised by Tricine-SDS-PAGE,at the same time,antibacterial activity was analysised by Agar diffusion method.As a result,Tricine-SDS-PAGE analysis showed that the molecular weight of 3.0 ku,and the expression products of antimicrobial peptides Cecropin AD have antibacterial activity.In this study,antimicrobial peptide Cecropin AD was expressed successfully in Pichia pastoris SMD1168 by constitutive expression method.Constitutive expression method was applied successfully for the future experimental study has laid a theoretical basis.
出处
《中国畜牧杂志》
CAS
北大核心
2010年第5期51-54,共4页
Chinese Journal of Animal Science
基金
北京市重大科技计划课题(D09010104270000)
