黏虫新型抗菌肽<i>Mysdiap</i>基因克隆、序列分析及原核表达

doi:10.7668/hbnxb.20193801

摘要/Abstract

摘要：

利用反转录PCR(RT-PCR)和末端快速扩增(RACE)技术从三龄黏虫体内获得新型抗菌肽Diapausin基因的全长序列,利用生物信息学软件对该序列进行分析,并用原核表达体系对其进行诱导表达,为探究该基因的结构和功能提供理论参考。结果表明,成功克隆到了黏虫新型抗菌肽Diapausin基因,命名为Mysdiap (GenBank登录号:AZJ51075)。该基因全长537 bp,其中5'端和3'端非编码区分别为63,279 bp,开放阅读框全长195 bp,编码64个氨基酸残基,预测蛋白质分子质量为7.13 ku,等电点为5.59。Mysdiap的N端前24个氨基酸残基为信号肽序列。多重联配结果表明,Mysdiap氨基酸序列中具有高度保守区域ECCRAHG。成功构建了pET-Mysdiap重组表达质粒,并在大肠杆菌中用IPTG诱导其表达,表达的重组蛋白分子质量在20 ku左右,以包涵体和可溶性蛋白形式存在。综上,从黏虫中克隆获得了新型抗菌肽基因Mysdiap的全长序列,并在大肠杆菌中成功诱导其表达。

关键词: 黏虫, 抗菌肽, Diapausin基因, 基因克隆, 原核表达

Abstract:

The full-length sequence of Diapausin,a novel antimicrobial peptide gene,was obtained from the third instar of oriental armyworm(Mythimna separata)by RT-PCR and RACE.And then,the amino acid sequence of Diapausin gene was analyzed by bioinformatics software.Finally,the prokaryotic expression system was used to induce the protein expression.This study would lay a theoretical foundation for further parsing of the function and structure of Diapausin gene in M.separata. Results showed that the novel antimicrobial peptide gene Diapausin was successfully cloned from M.separata and named Mysdiap (GenBank No.AZJ51075).The total length of this gene was 537 bp,with the 5' and 3' non-coding ends of 63,279 bp,respectively.The full length of the open reading frame was 195 bp,encoding 64 amino acid residues.The molecular weight and the isoelectric point of the protein were 7.13 ku and 5.59,respectively.The first 24 N-terminal amino acid residues of Mysdiap were signal peptide sequences.Multiple alignment of amino acid sequences showed that Mysdiap had a highly conserved region ECCRAHG.The recombinant expression plasmid pET-Mysdiap was successfully constructed and induced to express proteins by IPTG in Escherichia coli.The molecular weight of the expressed recombinant protein was about 20 ku,which could exist in both inclusion bodies and soluble proteins.In summary,the full-length sequence of the novel antimicrobial peptide gene Mysdiap was cloned from M.separata and successfully induced to express proteins in E.coli.

Key words: Mythimna separata, Antibacterial peptide, Diapausin gene, Gene cloning, Prokaryotic expression

张元臣, 胡梦杰, 薛爽, 郭文军, 沈红, 王景顺. 黏虫新型抗菌肽Mysdiap基因克隆、序列分析及原核表达[J]. 华北农学报, 2023, 38(6): 184-191. doi: 10.7668/hbnxb.20193801.

ZHANG Yuanchen, HU Mengjie, XUE Shuang, GUO Wenjun, SHEN Hong, WANG Jingshun. Cloning,Sequence Analysis and Prokaryotic Expression of Novel Antimicrobial Peptide Gene Mysdiap in Oriental Armyworm,Mythimna separata[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(6): 184-191. doi: 10.7668/hbnxb.20193801.

http://www.hbnxb.net/CN/Y2023/V38/I6/184

图/表 9

表1 本研究所用引物及用途

Tab.1 Primers used in this study and their purposes

引物名称 Primer name	引物序列(5'—3') Nucleotide sequence of primer(5'—3')	用途 Purpose
Diap-F1	ATGGCCGCCATCAAGACCGTTT	部分cDNA序列扩增
Diap-R1	GTAGCCGGAGTACCCATGAGC
Diap-5-Outer	CAGCTGCTGTAGCCGGAGTACCCA	5'端RACE
Diap-5-Inner	GCCATTGCAGGCTTCGACGCGGT
Diap-3-Inner	TGATGTTTACCGTGGAGGCAAACC	3'端RACE
Diap-F2	ATGGCCGCCATCAAGACCGTTT	全长序列验证
Diap-R2	TCAGTAACAAATCATTTGGCCG
Diap-F3	GAATTCAACCGCGTCGAAGCCTGCAAT	基因原核表达
Diap-R3	AAGCTTTCAGTAACAAATCATTTGGCC
M13-F	CAGGGTTTTCCCAGTCACG	菌液PCR
M13-R	GAGCGGATAACAATTTCACAC

图1 Mysdiap基因的PCR产物电泳结果 M.DL2000 DNA 分子质量标准;1,2.Mysdiap片段PCR产物;3,4.3'RACE 产物;5.5' RACE产物。

Fig.1 Electrophoretic result of PCR product from Mysdiap M.DL2000 DNA Marker;1,2.PCR product of partial CDS of Mysdiap;3,4.PCR product of Mysdiap via 3'RACE;5.PCR product of Mysdiap via 5'RACE.

图2 Mysdiap基因核苷酸序列及其翻译的氨基酸序列红色下划线.预测的信号肽序列;绿色下划线.Poly(A)加尾信号;实线方框.预测的磷酸化位点;*.终止密码子。

Fig.2 Nucleotide and translated amino acid sequences of Mysdiap The red underline.The sequence of predicted signal peptide;The green underline.Polyadenylation signal domain;Boxed in solid line.The predicted phosphorylation site;*.Stop codon.

图3 Mysdiap蛋白的保守结构域预测

Fig.3 Conserved domains prediction of Mysdiap protein

图4 Mysdiap与其他昆虫物种同源氨基酸的多重联配红线标注区域表示氨基酸序列的结构域。黑色、粉红色和蓝色阴影分别代表一致性为100%,≥75%和≥50%。Spldiap.斜纹夜蛾Diapausin蛋白 (登录号:ABU96713);SpediapA1.甜菜夜蛾Diapausin蛋白 (登录号:AKJ54510);SpediapA4.甜菜夜蛾Diapausin蛋白(登录号:AKJ54513);Osfdiap.亚洲玉米螟Diapausin蛋白(登录号:XP_028158203);SpediapB1.甜菜夜蛾Diapausin蛋白(登录号:AKJ54517);Mysdiap.黏虫Diapausin蛋白(登录号:AZJ51075);Legldiap.大豆食心虫Diapausin蛋白(登录号:XP_048002774);Masediap.烟草天蛾Diapausin蛋白(登录号:KAG6465316);Orcdiap.跳虫Diapausin蛋白(登录号:ODM87430)。

Fig.4 Multiple alignment based on amino acid sequences of Diapausin proteins from M.separata and other insects Area marked in red line is the domain of the amino acid sequence.The black,pink and blue shadows delegate the homology of amino acid at 100%,≥75% and ≥50% levels,respectively.Spldiap.Diapausin protein from Spodoptera litura (GenBank No.ABU96713);SpediapA1.Diapausin protein from S.exigua (GenBank No.AKJ54510);SpediapA4.Diapausin protein from S.exigua (GenBank No.AKJ54513);Osfdiap.Diapausin protein from Ostrinia furnacalis (GenBank No.XP_028158203);SpediapB1.Diapausin protein from S.exigua (GenBank No.AKJ54517); Mysdiap.Diapausin protein from Mythimna separata (GenBank No.AZJ51075);Legldiap.Diapausin protein from Leguminivora glycinivorella (GenBank No.XP_048002774);Masediap.Diapausin protein from Manduca sexta (GenBank No.KAG6465316);Orcdiap.Diapausin protein from Orchesella cincta(GenBank No.ODM87430).

图5 黏虫和其他昆虫Diapausin氨基酸序列的系统进化树

Fig.5 Phylogenetic tree analysis based on amino acid sequences of Diapausin proteins from M.separata and other insects

图6 重组表达质粒pET-Mysdiap的PCR扩增鉴定(A)和双酶切鉴定(B) M.DL2000 DNA分子质量标准;1,2.重组表达质粒PCR扩增产物;3,4.重组表达质粒的双酶切产物。

Fig.6 Identification of pET-Mysdiap plasmids by PCR(A)and double digestion(B) M.DL2000 DNA Marker;1,2.PCR product of pET-Mysdiap;3,4.Double digestion product of pET-Mysdiap.

图7 Mysdiap重组蛋白的SDS-PAGE分析 M.蛋白质分子质量标准;1.空质粒经1 μL IPTG诱导;2.空质粒经3 μL IPTG诱导;3.重组质粒经0 μL IPTG诱导;4.重组质粒经1 μL IPTG诱导;5.重组质粒经3 μL IPTG诱导。红色箭头指示目的蛋白。

Fig.7 SDS-PAGE analysis of the recombinant protein of Mysdiap M.Protein Marker;1.Induction of empty vector by 1 μL IPTG;2.Induction of empty vector by 3 μL IPTG;3.Induction of recombinant vector by 0 μL IPTG;4.Induction of recombinant vector by 1 μL IPTG;5.Induction of recombinant vector by 3 μL IPTG.The red arrows indicate the target protein.

图8 表达菌裂解液上清和沉淀中的Mysdiap重组蛋白检测 M.蛋白质分子质量标准;1.裂解沉淀;2.裂解上清。

Fig.8 Detection of Mysdiap protein in the supernatant and precipitate of lysis solution of expressing bacteria M.Protein Marker;1.The precipitate;2.The supernatant.

参考文献 34

[1]	江幸福, 张蕾, 程云霞, 罗礼智. 我国粘虫发生危害新特点及趋势分析[J]. 应用昆虫学报, 2014, 51(6):1444-1449.doi:10.7679/j.issn.2095-1353.2014.168.
	Jiang X F, Zhang L, Cheng Y X, Luo L Z. Novel features,occurrence trends and economic impact of the oriental armyworm,Mythimna separata(Walker)in China[J]. Chinese Journal of Applied Entomolog, 2014, 51(6):1444-1449.
[2]	段云, 陈琦, 郭培, 苗进, 夏鹏亮, 巩中军, 蒋月丽, 李彤, 武予清. 劳氏粘虫的发生危害和防治研究进展[J]. 昆虫学报, 2022, 65(4):522-532.doi:10.16380/j.kcxb.2022.04.012.
	Duan Y, Chen Q, Guo P, Miao J, Xia P L, Gong Z J, Jiang Y L, Li T, Wu Y Q. Research progress on the occurrence,damage and control of Mythimna loreyi(Lepidoptera:Noctuidae)[J]. Acta Entomologica Sinica, 2022, 65(4):522-532.
[3]	田晓丽, 卢文涛. 灭幼脲对粘虫围食膜的影响及与其病毒的联合毒力作用[J]. 西南农业学报, 2018, 31(1):89-93.doi:10.16213/j.cnki.scjas.2018.1.016.
	Tian X L, Lu W T. Effect of diflubenzuron on peritrophic membrane of Mythimna separata (walker)and allied toxicity function of diflubenzuron and Pseudaletia nuclear polyhedrosis virus,PsNVP against M.separata[J]. Southwest China Journal of Agricultural Sciences, 2018, 31(1):89-93.
[4]	曹庆军, 杨粉团, 梁尧, 姜晓莉, Lamine Diallo, 李刚. 灌浆期模拟粘虫危害对春玉米产量和品质的影响[J]. 华北农学报, 2015, 30(3):180-185.doi:10.7668/hbnxb.2015.03.031.
	Cao Q J, Yang F T, Liang Y, Jiang X L, Diallo L, Li G. Simulation in harm degree of Mythimna seperata in filling stage and its effect on maize yield and quality[J]. Acta Agriculturae Boreali-Sinica, 2015, 30(3):180-185.
[5]	Jiang X F, Luo L Z, Zhang L, Sappington T W, Hu Y. Regulation of migration in Mythimna separata (walker)in China:a review integrating environmental,physiological,hormonal,genetic,and molecular factors[J]. Environmental Entomolog, 2011, 40(3):516-533.doi:10.1603/EN10199. URL
[6]	张元臣, 蔡新, 张国强, 董丽丽, 王景顺. 黏虫溶菌酶基因Mswly的克隆与真核表达[J]. 河南农业科学, 2019, 48(5):70-77.doi:10.15933/j.cnki.1004-3268.2019.05.011.
	Zhang Y C, Cai X, Zhang G Q, Dong L L, Wang J S. Cloning and eukaryotic expression of Mswly in oriental armyworm,Mythimna separata[J]. Journal of Henan Agricultural Sciences, 2019, 48(5):70-77.
[7]	赵艳艳, 刘建兵, 罗梅浩, 郭线茹, 原国辉. 烟夜蛾组织蛋白酶B酶原基因的克隆、序列分析和原核表达[J]. 昆虫学报, 2008, 51(11):1121-1128.doi:10.3321/j.issn:0454-6296.2008.11.003.
	Zhao Y Y, Liu J B, Luo M H, Guo X R, Yuan G H. Cloning,sequence analysis and prokaryotic expression of cathepsin B gene from Helicoverpa assulta(Guene)(Lepidoptera:Noctuidae)[J]. Acta Entomologica Sinica, 2008, 51(11):1121-1128.
[8]	Charles H M, Killian K A. Response of the insect immune system to three different immune challenges[J]. Journal of Insect Physiolog, 2015, 81:97-108.doi:10.1016/j.jinsphys.2015.07.005.
[9]	Rolff J, Schmid Hempei P.Perspectives on the evolutionary ecology of arthropod antimicrobial peptides[J].Philosophical Transactions of the Royal Society of London Series B, Biological Sciences, 2016, 371(1695):20150297.doi:10.1098/rstb.2015.0297.
[10]	周琳. 雷公藤总生物碱对粘虫幼虫的毒力及对乙酰胆碱酯酶和乙酰胆碱的影响[J]. 华北农学报, 2011, 26(1):215-218.doi:10.7668/hbnxb.2011.01.044.
	Zhou L. Toxicity of total alkaloid from Tripterygium wilfordii hook against Mythimna separata(walker)and its effects of on AChE and Ach[J]. Acta Agriculturae Boreali-Sinica, 2011, 26(1):215-218.
[11]	Hillyer J F. Insect immunology and hematopoiesis[J]. Developmental & Comparative Immunolog, 2016, 58:102-118.doi:10.1016/j.dci.2015.12.006.
[12]	Lemaitre B, Hoffmann J. The host defense of Drosophila melanogaster[J]. Annual Review of Immunolog, 2007, 25(1):697-743.doi:10.1146/annurev.immunol.25.022106.141615.
[13]	Hultmark D, Steiner H, Rasmuson T, Boman H G. Insect immunity:purification and properties of three inducible bactericidal proteins from hemolymph of immunized pupae of Hyalophora cecropia[J]. European Journal of Biochemistr, 1980, 106(1):7-16.doi:10.1111/j.1432-1033.1980.tb05991.x.
[14]	Li Y M, Xiang Q, Zhang Q H, Huang Y D, Su Z J. Overview on the recent study of antimicrobial peptides:origins,functions,relative mechanisms and application[J]. Peptides, 2012, 37(2):207-215.doi:10.1016/j.peptides.2012.07.001. URL
[15]	Yi H Y, Chowdhury M, Huang Y D, Yu X Q. Insect antimicrobial peptides and their applications[J]. Applied Microbiology and Biotechnolog, 2014, 98(13):5807-5822.doi:10.1007/s00253-014-5792-6.
[16]	Eleftherianos I, Zhang W, Heryanto C, Mohamed A, Contreras G, Tettamanti G, Wink M, Bassal T. Diversity of insect antimicrobial peptides and proteins-a functional perspective:a review[J]. International Journal of Biological Macromolecules, 2021, 191:277-287.doi:10.1016/j.ijbiomac.2021.09.082. pmid: 34543628
[17]	Kouno T, Mizuguchi M, Tanaka H, Yang P, Mori Y, Shinoda H, Unoki K, Aizawa T, Demura M, Suzuki K, Kawano K. The structure of a novel insect peptide explains its Ca²⁺ channel blocking and antifungal activities[J]. Biochemistr, 2007, 46(48):13733-13741.doi:10.1021/bi701319t. URL
[18]	Sato K, Tanaka H, Saito Y, Suzuki K. Baculovirus-mediated production and antifungal activity of a diapause-specific peptide,diapausin,of the adult leaf beetle,Gastrophysa atrocyanea (Coleoptera:Chrysomelidae)[J]. Journal of Insect Biotechnology and Sericolog, 2002, 71(2):69-77.doi:10.11416/jibs2001.71.69.
[19]	He Y, Cao X L, Li K, Hu Y X, Chen Y R, Blissard G, Kanost M R, Jiang H B. A genome-wide analysis of antimicrobial effector genes and their transcription patterns in Manduca sexta[J]. Insect Biochemistry and Molecular Biolog, 2015, 62:23-37.doi:10.1016/j.ibmb.2015.01.015.
[20]	Wan H, Yuan M, You H, Li J H, Jin B R. Molecular cloning and characterization of a diapause-specific peptide in the beet armyworm,Spodoptera exigua[J]. Journal of Asia-Pacific Entomolog, 2012, 15(3):369-373.doi:10.1016/j.aspen.2012.02.009. URL
[21]	Al Souhail Q, Hiromasa Y, Rahnamaeian M, Giraldo M C, Takahashi D, Valent B, Vilcinskas A, Kanost M R. Characterization and regulation of expression of an antifungal peptide from hemolymph of an insect,Manduca sexta[J]. Developmental and Comparative Immunolog, 2016, 61:258-268.doi:10.1016/j.dci.2016.03.006.
[22]	Sanchez-Contreras M, Vlisidou I. The diversity of insect-bacteria interactions and its applications for disease control[J]. Biotechnology and Genetic Engineering Reviews, 2008, 25(1):203-243.doi:10.5661/bger-25-203. URL
[23]	Feng M, Fei S G, Xia J M, Labropoulou V, Swevers L, Sun J C. Antimicrobial peptides as potential antiviral factors in insect antiviral immune response[J]. Frontiers in Immunolog, 2020, 11:2030.doi:10.3389/fimmu.2020.02030.
[24]	Nunes C, Sucena É, Koyama T. Endocrine regulation of immunity in insects[J]. The FEBS Journal, 2021, 288(13):3928-3947.doi:10.1111/febs.15581. URL
[25]	Wang L, Qu L K, Lin S, Yang Q S, Zhang X X, Jin L B, Dong H, Sun D. Biological functions and applications of antimicrobial peptides[J]. Current Protein & Peptide Science, 2022, 23(4):226-247.doi:10.2174/1389203723666220519155942.
[26]	Flores-Villegas A L, Salazar-Schettino P M, Córdoba-Aguilar A, Gutiérrez-Cabrera A E, Rojas-Wastavino G E, Bucio-Torres M I, Cabrera-Bravo M. Immune defence mechanisms of triatomines against bacteria,viruses,fungi and parasites[J]. Bulletin of Entomological Research, 2015, 105(5):523-532.doi:10.1017/S0007485315000504. pmid: 26082354
[27]	Bao Y Y, Yamano Y, Morishima I. Induction of hemolin gene expression by bacterial cell wall components in eri-silkworm,Samia Cynthia ricini[J]. Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology, 2007, 146(1):147-151.doi:10.1016/j.cbpb.2006.10.092. URL
[28]	Levashina E A, Ohresser S, Lemaitre B, Imler J L. Two distinct pathways can control expression of the gene encoding the Drosophila antimicrobial peptide metchnikowin[J]. Journal of Molecular Biolog, 1998, 278(3):515-527.doi:10.1006/jmbi.1998.1705.
[29]	Mitpuangchon N, Nualcharoen K, Boonrotpong S, Engsontia P. identification of novel toxin genes from the stinging nettle caterpillar Parasa lepida (Cramer,1799):insights into the evolution of lepidoptera toxins[J]. Insects, 2021, 12(5):396.doi:10.3390/insects12050396. URL
[30]	Kim S Y, Chung H J, Thomas T L. Isolation of a novel class of bZIP transcription factors that interact with ABA-responsive and embryo-specification elements in the Dc3 promoter using a modified yeast one-hybrid system[J]. The Plant Journal:for Cell and Molecular Biology, 1997, 11(6):1237-1251.doi:10.1046/j.1365-313x.1997.11061237.x. URL
[31]	谢红艳, 万鲁长, 黄春燕, 刘国霞, Jean-Francois Picimbon, 宫志远. 烟粉虱MED隐种气味结合蛋白基因BtabOBP2和BtabOBP4的cDNA克隆及原核表达[J]. 环境昆虫学报, 2017, 39(4):752-761.doi:10.3969/j.issn.1674-0858.2017.04.3.
	Xie H Y, Wan L C, Huang C Y, Liu G X, Picimbon J F, Gong Z Y. cDNA cloning and prokaryotic expression of odorant binding protein genes BtabOBP2 and BtabOBP4 from the Bemisia tabaci MED cryptic species[J]. Journal of Environmental Entomolog, 2017, 39(4):752-761.
[32]	姚亚文, 赵其平, 朱顺海, 黄兵, 董辉, 黄文浩, 刘展, 于钰, 韩红玉. 柔嫩艾美耳球虫保守蛋白基因EtCHP40的分子特性研究[J]. 复旦学报(自然科学版), 2021, 60(1):102-110.
	Yao Y W, Zhao Q P, Zhu S H, Huang B, Dong H, Huang W H, Liu Z, Yu Y, Han H Y. Molercular characteristics of a conserved hypothetical protein gene EtCHP40 in Eimeria tenella[J]. Journal of Fudan University(Natural Science), 2021, 60(1):102-110.
[33]	唐威华, 张景六, 王宗阳, 洪孟民. SDS-PAGE 法测定His-tag融合蛋白分子量产生偏差的原因[J]. 植物生理学报, 2000, 26(1):64-68.doi:10.3321/j.issn:1671-3877.2000.01.012.
	Tang W H, Zhang J L, Wang Z Y, Hong M M. Causes of deviation made in determination of molecular weight of His-tag fusion protein by SDS-PAGE method[J]. Acta Phytophysiologica Sinica, 2000, 26(1):64-68.
[34]	Wang Z Q, Feng K, Tang F, Xu M. Activation of the host immune response in Hyphantria cunea (Drury)(Lepidoptera:Noctuidae)induced by Serratia marcescens Bizio[J]. Insects, 2021, 12(11):983.doi:10.3390/insects12110983. URL

选择文件类型/文献管理软件名称

选择包含的内容

黏虫新型抗菌肽Mysdiap基因克隆、序列分析及原核表达

Cloning,Sequence Analysis and Prokaryotic Expression of Novel Antimicrobial Peptide Gene Mysdiap in Oriental Armyworm,Mythimna separata

RichHTML

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 34

相关文章 15

编辑推荐

Metrics

本文评价

[1]	陈娜, 詹文文, 刘兴雨, 石磊鑫, 李若楠, 谢镕, 却志群. *番茄WRKY转录因子SlWRKY75的克隆、表达及功能分析*[J]. 华北农学报, 2023, 38(6): 1-10.
[2]	蔡诚诚, 李罗品, 温和, 刘石锋, 王强, 李立芹, 王西瑶. 马铃薯蔗糖磷酸合成酶StSPS1的原核表达及抗体制备[J]. 华北农学报, 2023, 38(6): 11-17.
[3]	柳婷, 李敏敏, 赖章龙, 刘彦斌, 刘凯, 肖伟, 赛清云, 田永华, 杨立强, 阮超岭, 杨瑞兰, 刘哲, 连总强. *兰州鲇Dmrta1基因克隆及其组织时空表达规律*[J]. 华北农学报, 2023, 38(6): 228-238.
[4]	袁惠君, 张瑞艳, 关玉晨, 余诗曼, 徐琰莹, 马倩国, 鲍婧婷. *扁果枸杞表皮蜡质转运相关基因LbABCG11的克隆及其表达特征分析*[J]. 华北农学报, 2023, 38(6): 45-54.
[5]	赖章龙, 阮超岭, 肖伟, 刘彦斌, 俞兆曦, 刘凯, 赛清云, 田永华, 张锋, 杨瑞兰, 柳婷, 杨立强, 王玉涛, 连总强. *兰州鲇NPY基因的克隆和表达分析*[J]. 华北农学报, 2023, 38(5): 227-238.
[6]	黄渺, 罗悠悠, 倪芮, 肇瑾. *观赏辣椒ARP基因克隆及表达分析*[J]. 华北农学报, 2023, 38(5): 69-76.
[7]	王茂存, 曹嘉伟, 周贺, 贾明轩, 魏淑珍, 巩校东, 刘玉卫, 谷守芹, 董金皋. *玉米大斑病菌转录因子基因StbZIP9的克隆、表达分析及其下游靶基因的筛选*[J]. 华北农学报, 2023, 38(5): 158-165.
[8]	杨宁宁, 徐明国, 张江伟, 易继海, 陈创夫. *牛病毒性腹泻病毒NS3基因的生物信息学分析、蛋白纯化及免疫原性分析*[J]. 华北农学报, 2023, 38(4): 205-212.
[9]	王可欣, 郭昭阳, 殷宇航, 陈胜忠, 宋希云, 赵美爱. *玉米ZmGST基因的克隆及表达特性分析*[J]. 华北农学报, 2023, 38(4): 20-27.
[10]	聂利珍, 张志成, 谢锐, 常悦, 张琼琳, 韩平安, 羿静. *彩色马铃薯StANS1a基因的克隆及表达分析*[J]. 华北农学报, 2023, 38(4): 38-46.
[11]	周倩怡, 黄思杰, 田洁. 大蒜2个中性/碱性转化酶基因AsNI的克隆与表达分析[J]. 华北农学报, 2023, 38(4): 54-64.
[12]	陈阳, 王琦, 高岩松, 尤学, 熊毅, 金一锋. *草地早熟禾蛋白激酶CIPK32基因克隆及非生物胁迫响应分析*[J]. 华北农学报, 2023, 38(4): 65-73.
[13]	赵胜男, 高美欣, 于朝杭, 聂凯悦, 王浩然, 孟杰, 朱虹, 李帅. *大豆GmFLK基因的克隆及特性分析*[J]. 华北农学报, 2023, 38(3): 26-34.
[14]	张沛沛, 陈涛, 景凡丽, 刘媛, 马靖福, 田甜, 王鹏, 杨德龙. *小麦TaPSKR1基因的克隆与表达分析*[J]. 华北农学报, 2023, 38(3): 1-9.
[15]	王鹏, 田哲娟, 赵雪芳, 康忱, 吴志明, 李亚栋, 黄冀楠. *番茄SlCaM3、SlCaM4和SlCaM5基因的克隆及低温胁迫下的表达分析*[J]. 华北农学报, 2023, 38(2): 75-84.

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

黏虫新型抗菌肽Mysdiap基因克隆、序列分析及原核表达

Cloning,Sequence Analysis and Prokaryotic Expression of Novel Antimicrobial Peptide Gene Mysdiap in Oriental Armyworm,Mythimna separata

RichHTML

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 34

相关文章 15

编辑推荐

Metrics

本文评价