【摘要】 【目的】为探讨烟蚜Myzus persicae适应UV-B胁迫的分子机制。【方法】采用RT-PCR和RACE技术克隆了烟蚜热激蛋白基因Hsp70的全长,利用生物信息学方法分析了其特征;采用实时荧光定量PCR检测了不同时长(0,15,30,60,90和120 min) UV-B胁迫下烟蚜成虫中该基因的相对表达量。【结果】克隆获得烟蚜Hsp70基因并命名为MpHsp70(GenBank登录号:MF509827),该基因全长为2 221 bp,开放阅读框(ORF) 1 965 bp,编码654个氨基酸,蛋白相对分子量为71. 41kD,等电点(p I)为5. 34,末端高度保守序列EEVD显示该蛋白属于胞质热激蛋白。系统进化关系分析表明,MpHsp70与多种昆虫的Hsp70的同源性较高,表现出Hsp70基因的高度保守性。实时荧光定量PCR分析表明,随着UV-B照射时间的延长,烟蚜成虫体内MpHsp70基因的表达量先上升后下降,当照射时间为30 min时表达量最大。【结论】烟蚜MpHsp70基因可以响应UV-B的胁迫,它可能在烟蚜适应UV-B胁迫过程中起到重要作用。更多还原

【Abstract】 【Aim 】 To explore the molecular mechanism of Myzus persicae to adapt to UV-B stress.【Methods】The full-length cDNA of the heat shock protein gene Hsp70 was cloned from M. persicae by using RT-PCR and RACE techniques,and its sequence characteristics were analyzed by bioinformatics methods. The relative expression levels of this gene in M. persicae adults exposed to UV-B stress for different time(0,15,30,60,90 and 120 min) were detected by qPCR. 【Results】The cDNA of a Hsp70 gene was cloned from M. persicae and named as MpHsp70(GenBank accession no. : MF509827).The full length of the gene is 2 221 bp and the open reading frame(ORF) is 1 965 bp,encoding 654 amino acids. The relative molecular weight of its encoded protein is 71. 41 k D,and the isoelectric point(pI) is 5. 34. The highly conserved sequence EEVD shows that this protein belongs to the cytoplasmic heat shock protein(Hsp). The phylogenetic relationship analysis showed that Mp Hsp70 is highly homologous to Hsp70 s of various insects and shows the high conservation characteristic of Hsp70 gene.The qPCR results showed that the relative expression level of MpHsp70 in M. persicae adults increased first,achieved the highest at 30 min post exposure,and then decreased with the prolonged UV-B irradiation time. 【Conclusion】MpHsp70 gene in M. persicae can respond to UV-B stress and may play an important role in the adaptation of M. persicae to UV-B stress.更多还原