小菜蛾磷酸丝氨酸磷酸酶基因的克隆与原核异源表达

Cloning and heterologous expression of Plutella xylostella L. phosphoserine phosphatase

【目的】克隆小菜蛾磷酸丝氨酸磷酸酶基因(Plutella xylostellaphosphoserine phosphatase,PxPP),并进行原核异源表达,为小菜蛾抗性机理研究奠定基础。【方法】提取小菜蛾的总RNA,反转录获得总cDNA,采用RT-PCR方法扩增目的基因,将其克隆进T载体并测序,然后将目的基因克隆到大肠杆菌表达载体pET-28a(+)中表达。【结果】目的基因序列分析结果表明,PxPP阅读框全长693 bp,编码230个氨基酸,预测编码蛋白分子质量和等电点分别为25.6 ku和5.82,表达的融合蛋白分子质量为26.9 ku。【结论】成功克隆和表达了小菜蛾磷酸丝氨酸磷酸酶基因。

【Objective】cDNA cloning and heterologous expression of Plutella xylostella phosphoserine phosphatase were presented.This paper provides basic information for Plutella xylostella resistance mechanism researches.【Method】cDNA of Plutella xylostella phosphoserine phosphatase was amplified by PCR and cloned into pGEM-T for sequence analysis.Then the target gene was subcloned into pET-28a(+)for prokaryotic expression.【Result】The length of target gene is 693 bp,coding 230 amino acids.The predicted MW and pI are 25...