摘要
酚氧化酶原(proPO)系统属于一种酶联级系统,在凡纳滨对虾抵抗病原菌侵袭过程中具有重要作用。Real-time PCR检测技术已经成为国际上检测基因表达通用的方法,能对低丰度mRNA水平进行定量,可在体外对细胞活性进行评估,选择该方法进行外界环境因子胁迫下的凡纳滨对虾(Litopenaeus vannamei)组织proPO表达分析。在实时荧光定量PCR中,标准曲线法是最为简单、准确的定量方法。为了快速、准确的定量proPO基因在逆境下的表达水平,构建了正确、可靠的proPO基因表达标准品质粒和标准曲线。结果表明,该法所构建的标准曲线能够保证数据的精确性,满足实时荧光定量试验的要求,完全可以作为proPO基因荧光定量检测的参照标准。
Prophenoloxidase（proPO）system is a kind of enzyme-level system and plays an important role during the invasion against pathogenic bacteria for Litopenaeus vannamei.Real-time PCR detection technology has become a common international approach to detect gene expression,especially the level of mRNA with low abundance.Therefore,the real-time fluorescent quantitative PCR method was established to quantify the transcript level of proPO gene response to the environmental stresses in Litopenaeus vannamei.The result demonstrated that the plasmid standard has a better sensitivity and reproducibility.It can be applied in the real-time PCR experiments.
出处
《生物技术通报》
CAS
CSCD
北大核心
2011年第2期131-135,共5页
Biotechnology Bulletin
基金
浙江省教育厅基金项目(Y200804766)
