[1]陈敬祥,程杰,林同.松墨天牛dynamin-1-like protein基因的鉴定及表达分析[J].江苏农业学报,2017,(03):524-532.[doi:doi:10.3969/j.issn.1000-4440.2017.03.007]
　CHEN Jing-xiang,CHENG Jie,LIN Tong.Identification and expression of dynamin-1-like protein gene from Monochamus alternatus[J].,2017,(03):524-532.[doi:doi:10.3969/j.issn.1000-4440.2017.03.007]
点击复制

松墨天牛dynamin-1-like protein基因的鉴定及表达分析()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年03期
页码:
524-532
栏目:
植物保护
出版日期:
2017-06-30

文章信息/Info

Title:
Identification and expression of dynamin-1-like protein gene from Monochamus alternatus
作者:
陈敬祥; 程杰; 林同
（华南农业大学林学与风景园林学院，广东广州510640）
Author(s):
CHEN Jing-xiang; CHENG Jie; LIN Tong
（College of Forestry and Landscape Architecture，South China Agricultural University，Guangzhou 510640，China）
关键词:
关键词:松墨天牛; dynamin-1-like protein; cDNA文库; RT-qPCR
Keywords:
Monochamus alternatus; dynamin-1-like protein gene; cDNA library; RT-qPCR
分类号:
S763.38
DOI:
doi:10.3969/j.issn.1000-4440.2017.03.007
文献标志码:
A
摘要:
为了探讨GTP酶超基因家族中dynamin-1-like protein基因在昆虫中的表达特性，以松墨天牛为研究对象，从已构建的cDNA文库中筛选到松墨天牛dynamin-1-like protein基因，命名为MaDLP1(GenBank:KU245763)。该序列长为2 133 bp，编码710个氨基酸。由此预测的蛋白质二级结构主要由α螺旋与无规则卷曲组成，其次是β片层与β转角；MaDLP1基因编码蛋白质，定位于细胞核。通过DNAMAN软件比对发现MaDLP1与赤拟谷盗的DLP1同源性最高，为82%，且存在3个保守酶域；用Clustal X和MEGA4.0构建系统发育树，显示松墨天牛与赤拟谷盗处在同一分支。RT-qPCR分析结果显示，MaDLP1在各虫态不间断表达，幼虫期在5龄幼虫中表达量最高，化蛹期间表达量先上升后下降，羽化期间表达量表现为先上升后下降，并在初羽化的成虫中表达量达到最大值；MaDLP1在幼虫和成虫头部表达量较高，且在幼虫脂肪体中表达最高；成虫的足、翅、触角和卵巢中也都有表达。说明MaDLP1的表达与松墨天牛的完全变态发育相关。
Abstract:
To characterize the expression of dynamin-1-like protein gene of GTPase supergene family in insects, the gene named MaDLP1 (GenBank: KU245763) was screened from the cDNA library of Monochamus alternatus. The sequence of MaDLP1 was 2133 bp in length, encoding 710 amino acids. The predicted secondary structure of MaDLP1 was mainly composed of alpha helix and random curl, followed by the beta turn and beta sheet. Subcellular localization of the gene was in nucleus. Amino acids sequence multiple alignment analysis of MaDLP1 showed the highest homology with Tribolium castaneum (82%) and there are three conservative enzyme domains. Phylogenetic tree built by Clustal X and MEGA4 indicated M. alternatus with T.castaneum were in the same branch. RT-qPCR revealed that MaDLP1 expression may be associated with holometabolous development of M. alternatus; expression level peaked in the fifth instar larvae at larval phase, fell after rose in larval-pupal and eclosion phases, and reached the maximum amount in pupa-adult. The expression level was higher in the heads of larvae and adults, and was the highest in fat body.

参考文献/References:

[1]张心团，赵和平，樊美珍，等. 松墨天牛生物学特性的研究进展[J . 安徽农业大学学报，2004,31(2):156-157.
[2]王文琪．赣北松墨天牛成虫发生及产卵规律［J］．江苏农业科学，2015,43(4):170-172.
[3]MAMIYA Y, ENDA N .Transmission of Bursaphelenchus lignicolus (Nematoda: Aphelenchoididae) Monochamus alternatus (Coleoptera: Cerambycidae)[J]. Nematologica , 1972, 18(2):159-162.
[4]蔡紫玲，陈敬祥，林同. 松墨天牛葡萄糖-6-磷酸异构酶基因克隆及序列分析[J]. 南方农业学报,2016，47(5)：657-661.
[5]ROMANI P, PAPI A, IGNESTI M,et al.Dynamin controls extracellular level of Awd/Nme1 metastasis suppressor protein[J].Naunyn-Schmiedeberg’s Archives of Pharmacology, 2016, 389(11):1-12.
[6]PRAEFCKE G J, MCMAHON H T. The dynamin superfamily: universal membrane tubulation and fission molecules?[J]. Nature Reviews Molecular Cell Biology, 2004, 5(2):133-147.
[7]MIYAGISHIMA S Y,KUWAYAMA H,URUSHIHARA H,et al. Evolutionary linkage between eukaryotic cytokinesis and chloroplast division by dynamin proteins[J]. Proceedings of the National Academy of Sciences, 2008, 105(39):15202-15207.
[8]WANG W,WANG X,FUJIOKA H,et al.Parkinson’s disease-associated mutant VPS35 causes mitochondrial dysfunction byrecycling DLP1 complexes[J].Nature Medicine, 2016, 22(1):54-63.
[9] WANG X,YAN M H,FUJIOKA H,et al.LRRK2 regulates mitochondrial dynamics and function through direct interaction with DLP1[J].Human Molecular Genetics, 2012, 21(9):1931-1944.
[10]WANG X,SU B,FUJIOKA H,et al.Dynamin-Like protein 1 reduction underlies mitochondrial morphology and distribution abnormalities in fibroblasts from sporadic alzheimer’s disease patients[J].American Journal of Pathology, 2008, 173(2):470-482.
[11]ITOYAMA A, MICHIYUKI S,HONSHO M,et al.Mff functions with Pex11pb and DLP1 in peroxisomal fission[J].Biology Open,2013, 2(10):998-1006.
[12]CHERRY S, PERRIMON N. Entry is a rate-limiting step for viral infection in a Drosophila melanogaster model of pathogenesis[J]. Nature Immunology, 2004, 5(1):81-87.
[13]MARSH M, HELENIUS A. Virus entry: open sesame[J].Cell, 2006, 124(4):729-740.
[14]NAKABACHI A, MIYAGISHIMA S, O′BROCHTA D,et al.Expansion of genes encoding a novel type of dynamin in the genome of the pea aphid, Acyrthosiphon pisum[J].Insect Molecular Biology, 2010, 19 (S2):165-173.
[15]MANALIL N S, JUNIOR TO V S, BRAITHWAITE K,et al.Comparative analysis of the Metarhizium anisopliae secretome in response to exposure to the greyback cane grub and grub cuticles[J].Fungal Biology,2010, 114(8):637-645.
[16]BARRIAS E S, CARVALHO T M U D, SOUZA W D.Trypanosoma cruzi: entry into mammalian host cells and parasitophorous vacuole formation[J].Frontiers in Immunology, 2013, 4:186.
[17]韦春梅，罗琳琳，吴华俊，等.松墨天牛幼虫cDNA文库的构建及EST分析[J].基因组学与应用生物学， 2014,33（1）：113-120.
[18]NIEMANN H H, KNETSCH M L W, SCHERER A, et al . Crystal structure of a dynamin GTPase domain in both nucleotide-free and GDP-bound forms[J].Embo Journal， 2001, 20(21):5813-5821.
[19]SARASTE M, SIBBALD P R, WITTINGHOFER A .The P-loop - a common motif in ATP- and GTP-binding proteins[J]. Trends in Biochemical Sciences, 1990, 15(11):430-434.
[20]VETTER I R, WITTINGHOFER A. The guanine Nucleotide-binding switch in three dimensions[J]. Science, 2001, 294(5545):1299-1304.
[21]BOCOS C, GOTTLICHER M, GEARING K,et al. Fatty acid activation of peroxisome proliferator-activated receptor(PPAR)[J].Journal of Steroid Biochemistry & Molecular Biology, 1995, 53(53):467-473.
[22]WANDERS R J, WATERHAM H R.Peroxisomal disorders: the single peroxisomal enzyme deficiencies.[J]. Biochimica Et Biophysica Acta, 2006, 1763(12):1707-1720.
[23]ISLINGER M, CARDOSO M J, SCHRADER M. Be different - the diversity of peroxisomes in the animal kingdom[J]. Biochimica Et Biophysica Acta, 2010, 1803(8):881-897.
[24]LI X,GOULD S J.The Dynamin-like GTPase DLP1 is essential for peroxisome division and is recruited to peroxisomes in part by PEX11*[J].Journal of Biological Chemistry, 2003, 278(19):17012-17020.
[25]KAHSAI L， MARTIN J R， WINTHER A M.Neuropeptides in the Drosophila central complex in modulation of locomotor behavior[J].Journal of Experimental Biology, 2010, 213(13):2256-2265.
[26]YOON Y,PITTS K R,MCNIVEN M A. Mammalian dynamin-like protein DLP1 tubulates membranes[J].Molecular Biology of the Cell, 2001, 12(9):2894-2905.
[27]KONOPKA C A, SCHLEEDE J B, SKOP A R,et al.Dynamin and cytokinesis[J].Traffic, 2006, 7(3):239-247.
[28]CHAN D C. Mitochondria: dynamic organelles in disease, aging, and development[J].Cell, 2006, 125(125):1241-1252.
[29]WANG X, SU B, LEE H G, et al, Impaired balance of mitochondrial fission and fusion in Alzheimer’s disease[J]. Journal of Neuroscience the Official Journal of the Society for Neuroscience, 2009, 29(28):9090-9103.
[30]MOCZEK A P.Pupal remodeling and the development and evolution of sexual dimorphism in horned beetles[J].American Naturalist, 2007, 168(6):711-729.
[31]WILLIAMS D W ,TRUMAN J W. Remodeling dendrites during insect metamorphosis[J] .Journal of Neurobiology,2005, 64(1):24-33.

备注/Memo

备注/Memo:
收稿日期：2016-11-25 基金项目：国家自然科学基金项目（31470653）；广东省自然科学基金项目（2015A030313416） 作者简介：陈敬祥（1993-），男，湖北荆州人，硕士研究生，研究方向为昆虫分子生物学。（Tel）13424049618;(E-mail)2441430459@qq.com 通讯作者：林同，(E-mail)lintong@scau.edu.cn
更新日期/Last Update: 2017-06-29