中文摘要：Argonaute (AGO)蛋白是RNA诱导沉默复合物中必不可少的催化成分，并在RNA干扰中发挥着核心作用。通过生物信息学和cDNA末端快速扩增(RACE)技术相结合，从灰飞虱中克隆和表征假定的AGO家族成员ls-AGO1和ls-AGO2。逆转录- qPCR (RT-qPCR)结果表明：ls-AGO1和ls-AGO2在灰飞虱的所有发育阶段均有表达，并以卵中的mRNA丰度最高。研究结果表明： AGO蛋白家族成员在灰飞虱的不同阶段均有表达，并响应各种胁迫。因此，可利用AGO蛋白家族通过控制相关基因的表达来调控灰飞虱的胁迫反应。
 
外文摘要：Argonaute (AGO) proteins are essential catalytic components of the RNA-induced silencing complex and play central roles in RNA interference. Using a combination of bioinformatics and rapid amplification of cDNA ends (RACE) methods, putative AGO subfamily members, ls-AGO1 and ls-AGO2, were cloned and characterized from the small brown planthopper, Laodelphax striatellus. The open reading frame (ORF) of ls-AGO1 is 2,820 bp long, encoding a putative protein of 939 amino acid residues, and ls-AGO2 contains an ORF of 2,490 bp, encoding 829 amino acid residues. The expected conserved PAZ and PIWI domains, and the conserved Asp-Asp-His (DDH) catalytic triad motif in the PIWI domain were observed in both ls-AGO1 and ls-AGO2. Reverse transcription-qPCR (RT-qPCR) results showed that both ls-AGO1 and ls-AGO2 were expressed in all developmental stages of L. striatellus with highest mRNA abundance in eggs. Expression of ls-AGO1 and ls-AGO2 was significantly decreased in adult insects in response to acquisition of rice black-streaked dwarf virus by second instar nymphs. mRNA expression of ls-AGO1 was significantly downregulated in response to low and high temperatures, but expression of ls-AGO2 was only affected by low temperature. ls-AGO1 and ls-AGO2 were initially downregulated when insects were transferred from rice to maize and to the wild grass Brachypodium distachyon, but expression showed partial or complete recovery 7 days after transfer. These results document that AGO subfamily members of L. striatellus are ubiquitously expressed at different developmental stages and respond to various stresses. Thus, AGO subfamily may act in regulating the stress-response of L. striatellus by controlling related gene expression.
外文关键词：Argonaute; gene silencing; Laodelphax striatellus; mRNA expression; RNA interference
作者：Zhou, Yan-Ru; Li, Lin-Ying; Li, Jun-Min; 等
作者单位：浙江农业科学院
期刊名称：ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY
期刊影响因子：1.357
出版年份：2016
出版刊次：1
点击下载：灰飞虱（半翅目：飞虱科）体内的ARGONAUTE蛋白家族基因研究