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.
摘要
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NlPLE25与piggyBac转座子之间的交叉识别。
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Acknowledgments
This work was supported by the Foundation of Guangzhou Science and Technology Key Project (No. 201904020041), China.
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Jun LYU designed and performed the experiment, wrote and edited the manuscript. Qin SU, Jinhui LIU, Lin CHEN, and Jiawei SUN participated in part of the experiment and edited the manuscript. Wenqing ZHANG designed and supervised the experiment, wrote and edited the manuscript. All authors have read and approved the final version of the manuscript, and therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Jun LYU, Qin SU, Jinhui LIU, Lin CHEN, Jiawei SUN, and Wenqing ZHANG declare that they have no conflict of interest.
All institutional and national guidelines for the care and use of laboratory animals were followed.
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Tables S1–S3; Figs. S1–S4
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Lyu, J., Su, Q., Liu, J. et al. Functional characterization of piggyBac-like elements from Nilaparvata lugens (Stål) (Hemiptera: Delphacidae). J. Zhejiang Univ. Sci. B 23, 515–527 (2022). https://doi.org/10.1631/jzus.B2101090
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DOI: https://doi.org/10.1631/jzus.B2101090