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利用RNA干扰技术在豌豆蚜(Acyrthosiphon pisum)中进行基因敲低。

Gene knockdown by RNAi in the pea aphid Acyrthosiphon pisum.

作者信息

Jaubert-Possamai Stéphanie, Le Trionnaire Gaël, Bonhomme Joël, Christophides Georges K, Rispe Claude, Tagu Denis

机构信息

INRA, Agrocampus Rennes, UMR 1099 BiO3P (Biology of Organisms and Populations applied to Plant Protection), F-35653 LE RHEU, France.

出版信息

BMC Biotechnol. 2007 Sep 28;7:63. doi: 10.1186/1472-6750-7-63.

Abstract

BACKGROUND

RNA interference (RNAi) is a powerful method to inhibit gene expression in a sequence specific manner.

RESULTS

Here, we described the development of RNAi by micro-injection of double-stranded RNA (dsRNA) in the pea aphid Acyrthosiphon pisum. Injection of dsRNA into whole aphid body induced the silencing of two marker genes with different expression patterns: the ubiquitously expressed Ap-crt genes encoding a calreticulin and the gut specific Ap-cath-L gene encoding a cathepsin-L. Time-course analysis of the silencing showed similar temporal patterns for both genes: inhibition started at 1 day after injection, reached its maximum at 5 days and stopped at 7 days. A comparable 40% decrease of gene expression was observed for Ap-crt and Ap-cath-L.

CONCLUSION

The pea aphid is the first Hemipteran insect for which genome sequence will be available soon. The gene knockdown technique developed in this study will be an essential post-genomic tool for further investigations in aphidology.

摘要

背景

RNA干扰(RNAi)是一种以序列特异性方式抑制基因表达的强大方法。

结果

在此,我们描述了通过向豌豆蚜(Acyrthosiphon pisum)显微注射双链RNA(dsRNA)来开发RNA干扰技术的过程。将dsRNA注射到整个蚜虫体内可诱导两个具有不同表达模式的标记基因沉默:普遍表达的编码钙网蛋白的Ap-crt基因和肠道特异性的编码组织蛋白酶L的Ap-cath-L基因。对沉默的时间进程分析显示这两个基因具有相似的时间模式:抑制在注射后1天开始,在5天达到最大值,并在7天停止。观察到Ap-crt和Ap-cath-L的基因表达下降了约40%。

结论

豌豆蚜是第一种即将获得基因组序列的半翅目昆虫。本研究中开发的基因敲低技术将成为蚜虫学进一步研究中必不可少的后基因组工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbe9/2075495/50762ebe9e2f/1472-6750-7-63-1.jpg

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