Science and Technology Production
Congress
Authorship
Date
2015
Publishing House and Editing Place
BIOCELL
Summary
Information provided by the agent in
SIGEVA
Functional genomics is a field of life sciences which aims include the study of gene function in the context of an organism. Generally, these analyses require the introduction of mutations such as gene knock-outs with the goal to collaborate in the understanding of the role played by proteins and non-coding regions. For this purpose, the use of insertionaltransgenesis based on engineering transposons has been reported as a good molecular tool. PiggyBac, a class II mobile element isolated from i...
Functional genomics is a field of life sciences which aims include the study of gene function in the context of an organism. Generally, these analyses require the introduction of mutations such as gene knock-outs with the goal to collaborate in the understanding of the role played by proteins and non-coding regions. For this purpose, the use of insertionaltransgenesis based on engineering transposons has been reported as a good molecular tool. PiggyBac, a class II mobile element isolated from insects, is one of the most employed transposons in functional genomics having successful references in organisms from different lineages including bacteria and mammals. Baculoviruses are insect pathogens carrying large circular dsDNA genomes which are infective per se. They are very exploited in different technologies including protein expression, bioinsecticide uses and gene delivery in mammals, among others. Considering that the biological role of baculoviral gene content is not fully understood, we decided to develop an in vitro mutagenesis approach based on a binary system containing the recombinant PiggyBac transposase and an insertional cassette which confers a bacterial ORI to transform virus genomes in plasmids. In this work the heterologous expression of transposase in bacteria, Pichia pastoris and insect cells is reported and the preliminary activity studies are discussed.
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Key Words
TRANSPOSONPIGGYBACBACULOVIRUS