Construction of a cDNA library of Aphis gossypii Glover for use in RNAi

Aphis gossypii Glover is an important insect pest that functions as a viral vector and mediates approximately 45 different viral diseases. As part of a strategy for control of A. gossypii, we investigated the functions of genes using RNAi. To this end, a cDNA library was constructed for various gene...

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Bibliographic Details
Published in:Entomological research 2018, 48(5), , pp.384-389
Main Authors: Kwon, HyeRi, Kim, JungGyu, Lim, HyounSub, Yu, YongMan, Youn, YoungNam
Format: Article
Language:English
Subjects:
Aphis gossypii
cDNA library
Cloning
Colon
Gateway cloning system
Gene silencing
Genes
Mammals
Recombination
RNA-mediated interference
RNAi
Transfection
Vectors
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Summary:Aphis gossypii Glover is an important insect pest that functions as a viral vector and mediates approximately 45 different viral diseases. As part of a strategy for control of A. gossypii, we investigated the functions of genes using RNAi. To this end, a cDNA library was constructed for various genes and for selecting appropriate targets for RNAi mediated silencing. The cDNA library was constructed using the Gateway cloning system with site‐specific recombination of bacteriophage λ. It was used to carry out single step cloning of A. gossypii cDNAs. As a result, a cDNA library with a titer of 8.4 × 106 was constructed. Since the sequences in this library carry att sites, they can be cloned into various binary vectors. This library will be of value for various studies. For later screening of selected genes, it is planned to clone the library into virus‐induced gene silencing (VIGS) vectors, which makes it possible to analyze gene function and allow subsequent transfection of plants. Such transfection experiments will allow testing of RNAi‐induced insecticidal activity or repellent activity to A. gossypii, and result in the identification of target genes. It is also expected that the constructed cDNA library will be useful for analysis of gene functions in A. gossypii.
ISSN:1738-2297
1748-5967
1748-5967
DOI:10.1111/1748-5967.12301