The hAT-family transposable element, hopper, from Bactrocera dorsalis is a functional vector for insect germline transformation.

IF 2.9 Q2 Biochemistry, Genetics and Molecular Biology
Alfred M Handler, Marc F Schetelig
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引用次数: 1

Abstract

Background: The hopper hAT-family transposable element isolated from the Oriental fruit fly, Bactrocera dorsalis, is distantly related to both the Drosophila hobo element and the Activator element from maize. The original 3120 bp hopperBd-Kah element isolated from the Kahuku wild-type strain was highly degenerate and appeared to have a mutated transposase and terminal sequences, while a second 3131 bp element, hopperBd-we, isolated from a white eye mutant strain had an intact transposase reading frame and terminal sequences consistent with function.

Results: The hopperBd-we element was tested for function by its ability to mediate germline transformation in two dipteran species other than B. dorsalis. This was achieved by creating a binary vector/helper transformation system by linking the hopperBd-we transposase reading frame to a D. melanogaster hsp70 promoter for a heat-inducible transposase helper plasmid, and creating vectors marked with the D. melanogaster mini-white+ or polyubiquitin-regulated DsRed fluorescent protein markers.

Conclusions: Both vectors were successfully used to transform D. melanogaster, and the DsRed vector was also used to transform the Caribbean fruit fly, Anastrepha suspensa, indicating a wide range of hopper function in dipteran species and, potentially, non-dipteran species. This vector provides a new tool for insect genetic modification for both functional genomic analysis and the control of insect populations.

Abstract Image

Abstract Image

背小实蝇hat家族转座因子hopper是昆虫种系转化的功能载体。
背景:从东方果蝇小实蝇(Bactrocera dorsalis)中分离到的hopper hAT-family转座因子与来自玉米的Drosophila hobo元件和Activator元件均有远亲关系。从Kahuku野生型菌株中分离到的3120 bp的hopperBd-Kah元件高度退化,出现了转座酶和末端序列的突变,而从白眼突变株中分离到的3131 bp的hopperBd-we元件有完整的转座酶阅读框和与功能一致的末端序列。结果:hopperb -we元件在两种双翅目昆虫中介导种系转化的能力得到验证。这是通过建立一个二元载体/辅助转化系统实现的,通过将hopperBd-we转座酶阅读框连接到D. melanogaster热诱导转座酶辅助质粒的hsp70启动子,并创建带有D. melanogaster微白+或多泛素调节的DsRed荧光蛋白标记的载体。结论:两种媒介均成功转化了黑腹扁蝇,并且DsRed媒介也被用于转化加勒比果蝇,表明在双翅目和非双翅目物种中具有广泛的跳跃功能。该载体为昆虫功能基因组分析和昆虫种群控制提供了一种新的基因改造工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
BMC Genetics
BMC Genetics 生物-遗传学
CiteScore
4.30
自引率
0.00%
发文量
77
审稿时长
4-8 weeks
期刊介绍: BMC Genetics is an open access, peer-reviewed journal that considers articles on all aspects of inheritance and variation in individuals and among populations.
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