Cloning of Sodium Channel Genes of Drosophila simulans and Its Bioinformatics Analysis

doi:10.16409/j.cnki.2095-039x.2021.03.012

Abstract

Abstract: Voltage-gated sodium channel gene of Drosophila simulans was cloned using RT-PCR and typical characteristics and phylogenetic relationship of the gene were clarified. The open reading frame of the gene (GenBank number of MT113357) was 6270 bp in length and encoded 2090 amino acids. Bioinformatics analysis showed a 96.86% similarity of the sodium channel genes of D. simulans and Drosophila melanogaster, a 88.75% similarity of the sodium channel genes of D. simulans and Musca domestica, and a very high degree of similarity in the sodium channel genes of D. simulans and Diptera insects. The cloned sequence was of the basic characteristics of insect sodium channels, with a DEKA module and a hydrophobic inactivation gate MFM. Three alternative splicing exons i, a, and b and 17 RNA editings were obtained. The present cloning and analysis of the sodium channel gene of D. simulans serve to provide a foundation for resistance mechanism study and new ideas for the development of new agricultural pesticides.

Key words: Drosophila simulans, insect sodium channel, cloning, alternative splicing, bioinformatics analysis

CLC Number:

S476

DENG Denghui, DUAN Wenbo, WANG Hao, LI Fen, WU Shaoying. Cloning of Sodium Channel Genes of Drosophila simulans and Its Bioinformatics Analysis[J]. Chinese Journal of Biological Control, 2021, 37(2): 340-348.

References

[1] 薛万琦, 赵建铭. 中国蝇类(上册)[M]. 沈阳:辽宁科学技术出版社, 1996, 286-288, 294-295.
[2] 李群松. 杨梅红跟果蝇的发生及防治[J]. 农村实用技术, 2003(2):29.
[3] Komai T. Collection of D. simulans from Japan[J]. Drosophila Information Service, 1937, 8:78.
[4] Okada T. Systematic Study of Drosophilidae and Allied Families of Japan[M]. Tokyo:Gihodo, 1956.
[5] 钱远槐, 张菁, 曾庆韬, 等. 武汉地区果蝇调查研究(双翅目:果蝇科)[J]. 湖北大学学报(自然科学版), 1993, 15(3):315- 321.
[6] 徐书华, 曾庆韬, 钱远槐, 等. 中国大陆拟果蝇(Drosophila simulans)的随机扩增多态性DNA分析[J]. 遗传学报, 2003, 30(7):673-680.
[7] 太一梅, 朱斌, 何绍华, 等. 杨梅果蝇综合防治技术研究[J]. 植物保护, 2015, 41(1):193-195.
[8] 李学斌, 王林. 艾绿士防治杨梅果蝇药效试验[J]. 中国南方果树, 2012, 41(1):63-64.
[9] Jin T, Zeng L, Lin Y Y, et al. Insecticide resistance of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera:Tephritidae), in mainland China[J]. Pest Management Science, 2011, 67(3):370-376.
[10] 章与苹, 曾玲, 陆永跃, 等. 华南地区桔小实蝇抗药性动态监测[J]. 华南农业大学学报, 2007, 28(3):20-23.
[11] Catterall W. From ionic currents to molecular mechanisms:the structure and function of voltage-gated sodium channels[J]. Neuron 2000, 26(1):13-25.
[12] Dong K, Du Y Z, Rinkevich F, et al. Molecular biology of insect sodium channels and pyrethroid resistance[J]. Insect Biochemistry and Molecular Biology, 2014, 50(1):1-17.
[13] Dong K. A single amino acid change in the para sodium channel protein is associated with knockdown-resistance (kdr) to pyrethroid insecticides in German cockroach[J]. Insect Biochemistry and Molecular Biology, 1997, 27:93-100.
[14] Williamson M S, Martinez D, Torres C, et a1. Identification of mutations in the housefly para-type sodium channel gene associated with knockdown resistance (kdr) to pyrethroid insecticides[J]. Molecular and General Genetics,1996, 252:51-60.
[15] Martinez-ToITes D F, Chandre M S, Williamson, et a1. Molecular characterization of pyrethroid knockdown resistance (kdr) in the major malaria vector Anopheles gambiaess[J]. Insect Molecular Biology, 1998, 7:179-184.
[16] Park Y, Taylor R, Feyereisen R. A valine 421 to methionine mutation in IS6 of thehscp voltage-gated sodium channel associated with pyrethroid resistance in Heliothis virescens F[J]. Biochemistry and Biophysics,1997, 239:688-691.
[17] 曹晓梅, 孙晨熹. 钠离子通道与拟除虫菊酯击倒抗性的研究进展[J]. 中国媒介生物学及控制, 2001, 12(6):466-469.
[18] David P, Jacques B, Marcel A, et al. Pyrethroid receptor in the insect sodium channel:alteration of its properties in pyrethroid-resistant flies[J]. Biochemistry, 1989, 28(4):1673-1677.
[19] Tan J, Liu Z, Nomura Y, et al. Alternative splicing of an insect sodium channel gene generates pharmacologically distinct sodium channels[J]. Journal of Neuroscience, 2002, 22(13):5300-5309.
[20] 朱玉贤. 现代分子生物学[M]. 北京:中国高等教育出版社, 1997.
[21] Guindon S, Dufayard J F, Lefort V, et al. New algorithms and methods to estimate maximumlikelihood phylogenies:assessing the performance of phyML 3.0[J]. Systematic Biology, 2010, 59(3):307-321.
[22] Salkoff L, Butler A, Wei A, et al. Genomic organization and deduced amino acid sequence of a putative sodium channel gene in Drosophila[J]. Science 1987, 237(4816):744-749.
[23] Feng G, Deak P, Chopra M, et a1. Cloning and functional analysis of tipE, a novel membran e protein that enhances Drosophila para sodium channel function[J]. Cell, 1995, 82:1001- 1011.
[24] Goldin A L. Resurgence of sodium channel research[J]. Annual Review of Physio1ogy, 2001, 63:871-894.
[25] 王颢, 段文波, 李芬, 等. 麦长管蚜电压门控钠离子通道基因克隆及序列分析[J]. 中国生物防治学报, 2020, 36(2):307-314.
[26] Zhang K, Chen M L, Wang H, et al. Molecular characterization and functional expression of voltage-gated sodium channel variants in Apolygus lucorum (Meyer-Dür)[J]. Pest Management Science, 2020, 76(6):2095-2104.
[27] Thackeray J R, Ganetzky B. Conserved alternative splicing patterns and splicing signals in the Drosophila sodium channel gene para[J]. Genetics, 1995, 141(1):203-214.
[28] Juan J S, Jeffrey G S. Conservation of the voltage-sensitive sodium channel protein within the Insecta[J]. Insect Molecular Biology, 2019, 29(1):9-18.
[29] Song W, Liu Z, Tan J, et al. RNA editing generates tissue-specific sodium channels with distinct gating properties[J]. The Journal of Biological Chemistry, 2004, 279(31):32554-32561.
[30] Du Y, Lee J E, Nomura Y, et al. Identification of a cluster of residues in transmembrane segment 6 of domain Ⅲ of the cockroach sodium channel essential for the action of pyrethroid insecticides[J]. Biochemical Journal, 2009, 419(2):377-385.
[31] Lin W H, Wright D E, Muraro N I, et al. Alternative splicing in the voltage-gated sodium channel dmnav regulates activation, inactivation, and persistent current[J]. Journal of Neurophysiology, 2009, 102(3):1994-2006.
[32] Tan J, Liu Z, Nomura Y, Goldin A L, et al. Alternative splicing of an insect sodium channel gene generates pharmacologically distinct sodium channels[J]. Journal Neuroscience, 2002, 22(13):5300-5309.
[33] Lee S H, Ingles P J, Knipple D C, et al. Developmental regulation of alternative exon usage in the house fly vssc1sodium channel gene[J]. Invertebrate Neuroscience, 2002, 4(3):125-133.
[34] Chang C, Shen W K, Wang T T, et al. A novel amino acid substitution in a voltage-gated sodium channel is associated with knockdown resistance to permethrin in aedes aegypti[J]. Insect Biochemistry and Molecular Biology, 2009, 39(4):272-278.
[35] Rieder L E, Staber C J, Hoopengardner B, et al. Tertiary structural elements determine the extent and specificity of messenger RNA editing[J]. Nature Communications, 2013, 4:2232-3232.
[36] Hanrahan C J, Palladino M J, Ganetzky B, et al. RNA editing of the Drosophila para Nat channel transcript[J]. Genetics, 2000, 155(3):1149-1160.
[37] Reenan R A, Hanrahan C J, Ganetzky, B. The mle (napts) RNA helicase mutation in drosophila results in a splicing catastrophe of the para Nat channel transcript in a region of RNA editing[J]. Neuron, 2000, 25(1):139-149.
[38] Palladino M J, Keegan L P, O'Connell M A, et al. A-to-I pre-mRNA editing in Drosophila is primarily involved in adult nervous system function and integrity[J]. Cell, 2000, 102(4):437-449.
[39] Rieder L E, Staber C J, Hoopengardner B, et al. Tertiary structural elements determine the extent and specificity of messenger RNA editing[J]. Nature Communications, 2013, 4(4):1-11.
[40] Hopkins B W, Pietrantonio P V. The Helicoverpa zea (Boddie) (Lepidoptera:Noctuidae) voltage-gated sodium channel and mutations associated with pyrethroid resistance in field-collected adult males[J]. Insect Biochemistry and Molecular Biology, 2010, 40:385-393.
[41] Davies T G, Field L M, Usherwood P N, et al. A comparative study of voltage-gated sodium channels in the Insecta:implications for pyrethroid resistance in Anopheline and other Neopteran species[J]. Insect Molecular Biology, 2007, 16(3):361-375.
[42] Lee S H, Soderlund D M. The V410M mutation associated with pyrethroid resistance in Heliothis virescens reduces the pyrothroidse nsitivity of house fly sodium channels expressed in Xenopus oocytes[J]. Insect Molecular Biology, 2001, 31:19-29.
[43] Hu Z, Du Y, Nomura Y, et al. A sodium channel mutation identified in Aedes aegypti selectively reduces cockroach sodium channel sensitivity to type I, but not type Ⅱ pyrethroids[J]. Insect Molecular Biology, 2011, 41(1):9-13.