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Journal of Zhejiang University SCIENCE B

ISSN 1673-1581(Print), 1862-1783(Online), Monthly

Functional characterization of piggyBac-like elements from Nilaparvata lugens (Stål) (Hemiptera: Delphacidae)

Abstract: PiggyBac is a transposable DNA element originally discovered in the cabbage looper moth (Trichoplusia ni). The T. ni piggyBac transposon can introduce exogenous fragments into a genome, constructing a transgenic organism. Nevertheless, the comprehensive analysis of endogenous piggyBac-like elements (PLEs) is important before using piggyBac, because they may influence the genetic stability of transgenic lines. Herein, we conducted a genome-wide analysis of PLEs in the brown planthopper (BPH) Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), and identified a total of 28 PLE sequences. All N. lugens piggyBac-like elements (NlPLEs) were present as multiple copies in the genome of BPH. Among the identified NlPLEs, NlPLE25 had the highest copy number and it was distributed on five chromosomes. The full length of NlPLE25 consisted of terminal inverted repeats and sub-terminal inverted repeats at both terminals, as well as a single open reading frame transposase encoding 546 amino acids. Furthermore, NlPLE25 transposase caused precise excision and transposition in cultured insect cells and also restored the original TTAA target sequence after excision. A cross-recognition between the NlPLE25 transposon and the piggyBac transposon was also revealed in this study. These findings provide useful information for the construction of transgenic insect lines.

Key words: Nilaparvata lugens; piggyBac-like elements; Transposon; NlPLE25

Chinese Summary  <66> 褐飞虱中piggyBac类似元素的功能特征

吕军,苏秦,刘锦辉,陈麟,孙嘉伟,张文庆
有害生物控制与资源利用国家重点实验室/中山大学生命科学学院,中国广州市,510275
目的:鉴定褐飞虱中的piggyBac类似元素并评估其转座活性。
创新点:首次鉴定了褐飞虱中的piggyBac类似元素并发现NlPLE25具备转座活性。
方法:使用RepeatMasker和RepeatModeler软件预测褐飞虱基因组中的piggyBac类似元素序列。随后在S2细胞中测试了NlPLE25的切割活性,在Sf9细胞中测试了NlPLE25的转座能力。
结论:我们从褐飞虱基因组中鉴定出28个piggyBac类似元素。进一步分析NlPLE25发现其在昆虫细胞中引起精确的切除和转座。此外,我们还发现了NlPLE25piggyBac转座子之间的交叉识别。

关键词组:褐飞虱;piggyBac类似元素;转座子;NlPlE25


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DOI:

10.1631/jzus.B2101090

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On-line Access:

2022-06-08

Received:

2021-12-26

Revision Accepted:

2022-03-03

Crosschecked:

2022-06-08

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