中国水稻科学
     Home | About Journal | Editorial Board | Publication Ethics Statement | Subscriptions | Advertisement | Contacts Us | Chinese
  2010, Vol. 24 Issue (4): 353-359     DOI: 10.3969/j.issn.1001-7216.2010.04.04
研究报告 Current Issue | Next Issue | Archive | Adv Search  |   
Cloning, Expression and Sequence Analysis of G Protein β Subunit Gene of Rice False Smut Pathogen Ustilaginoidea virens
LIU Lian-meng, WANG Ling, HUANG Wen-wen, LIU En-yong, HUANG Shi-wen*
China National Rice Research Institute, Hangzhou 310006, China; *Corresponding author, E-mail: swhuang666@sohu.com
 Download: PDF (2155 KB)   HTML (1 KB)   Export: BibTeX | EndNote (RIS)      Supporting Info
Abstract Ustilagionidea virens is a common rice pathogen, which causes rice false smut. G protein may involve in the pathogenicity of U. virens. To study the role of G protein of U. virens in the pathogenicity, a gene that encodes G protein βsubunit had been isolated and characterized. The degenerate primers based on conserved domains of other reported genes were designed to obtain a homologous fragment. Thermal asymmetric interlaced PCR (TAILPCR) was used to isolate flank sequence of the homologous fragment. The gene was obtained by jointing all the fragments and named UvGβ1. Full length of UvGβ1 was 2037 bp, which contained 4 introns, 5 extrons, and an open reading frame (ORF) encoding 359 amino acids. The cDNA sequence containing entire ORF of UvGβ1 was cloned through RTPCR by primers based on the sequence of UvGβ1. The DNA and cDNA sequences of UvGβ1 have been registered in GenBank with accession numbers GU014921 and GU065745. Phylogenetic analysis showed that UvGβ1 had the nearest relationship with the G protein βsubunit gene of Cryphonectria parasitica. The ORF of UvGβ1 was subcloned into pET30a and was expressed in E. coli BL21. The recombinant protein was obtained by IPTG inducing.
Service
E-mail this article
Add to my bookshelf
Add to citation manager
E-mail Alert
RSS
Articles by authors
LIU Lian-meng
WANG Ling
HUANG Wen-wen
LIU En-yong
HUANG Shi-wen*
Key wordsUstilaginoidea virens   heterotrimeric G protein   G protein beta subunit gene   thermal asymmetric interlaced PCR   prokaryotic expression     
Received: 1900-01-01;
Cite this article:   
LIU Lian-meng,WANG Ling,HUANG Wen-wen et al. Cloning, Expression and Sequence Analysis of G Protein β Subunit Gene of Rice False Smut Pathogen Ustilaginoidea virens[J]. , 2010, 24(4): 353-359 .
 
[1] Koiso Y, Li Y, Iwasaki S, et al. Ustiloxins, antimitotic cyclic peptides from false smut balls on rice panicles caused by Ustilaginoidea virens. J Antibiot (Tokyo), 1994, 47(7): 765-773.
[2] Rush M C, Shahjahan A K M, Jones J P, et al. Outbreak of false smut of rice in Louisiana. Plant Dis, 2000, 84: 100.
[3] Gilbert R D, Dean R A. Chemical signals responsible for appressorium formation in rice blast fungus Magnaporthe grisea. Physiol & Mol Plant Pathol, 1996, 48: 335-346.
[4] 郝志敏, 李志勇, 董金皋, 等. 玉米大斑病菌G蛋白β亚基编码基因的克隆与表达. 农业生物技术学报, 2008, 16(6): 1025-1030.
[5] Blker M. Sex and crime: Heterotrimeric G proteins in fungal mating and pathogenesis. Fungal Genet Biol, 1998, 25: 143-156.
[6] Lengeler K B, Davidson R C, D’souza C, et al. Signal transduction cascades regulating fungal development and virulence. Microbiol Mol Biol Rev, 2000, 64: 746-785.
[7] Vanti W B, Swaminathan S, Blevins R, et al. Patent status of the therapeutically important G-protein-coupled receptors. Exp Opin Ther Patents, 2001, 11: 1861-1887.
[8] 吴志红, 汪天虹, 黄卫, 等. 简单易行的丝状真菌染色体DNA提取法.菌物系统, 2001, 20(4): 575-577.
[9] Liu Y G, Huang N. Efficient amplification of insert end sequences from bacterial artificial chromosome clones by thermal asymmetric interlaced PCR. Plant Mol Biol Rep, 1998, 16: 175-181.
[10] Gilman A G. G proteins: Transducers of receptor-generated signals. Annu Rev Biochem, 1987, 56: 615-649.
[11] Nishimura M, Park G, Xu G R. The G-beta subunit MGB1 is involved in regulating multiple steps of infection-related morphogenesis in Magnaporthe grisea. Mol Microbiol, 2003, 50: 231-243.
[12] Ganem S, Lu S W, Lee B N, et al. The G protein β subunit of Cochliobolus heterostrophus involved in virulence, asexual and sexual reproductive ability andmorphogenesis. Eukaryot Cell, 2004, 3: 1652-1663.
[13] Jain S, Akiyama K, Kan T, et al. The G protein β subunit FGB1 regulates development and pathogenicity in Fusarium oxysporum. Curr Genet, 2003, 43: 79-86.
[14] Yu H Y, Seo J A, Kim J E, et al. Functional analyses of heterotrimeric G protein Gα and Gβ subunits in Gibberella zeae. Microbiology, 2008, 154: 392-401.
[15] Kasahara S, Nuss D L. Targeted disruption of a fungal G protein β subunit gene results in increased vegetative growth but reduced virulence. Mol Plant Microbe Interact, 1997, 10(8): 984-993.
[16] Muller P, Leibbrandt A, Teuissen S, et al. The G β-subunit-encoding gene bpp1 controls cyclic-AMP signaling in Ustilago maydis. Eukaryot Cell, 2004, 3: 806-814.
[17] Sagaram U S, Shim W B. Fusarium verticillioides GBB1, a gene encoding heterotrimeric G protein β subunit, is associated with fumonisin B1 biosynthesis and hyphal development but not with fungal virulence. Mol Plant Pathol, 2007, 8: 375-384.
[1] WANG Ling,LIU Lian-meng,FU Qiang,HUANG Shi-wen*. Prokaryotic Expression, Purification and Activity Assay of Neomycin Phosphotransferase Ⅱ (nptⅡ) Gene[J]. , 2011, 25(3): 326-330 .
[2] CHEN Xi-jun ,LIU Xiao-wei ,ZUO Shi-min ,TONG Yun-hui ,PAN Xue-biao ,XU Jing-you . Prokaryotic Expression of Polygalacturonase-Inhibiting Protein Gene (Ospgip1) from Rice and Bioinformatics Analysis of Its Coding Product

[J]. , 2011, 25(2): 136-142 .
[3] OUYANG Yuan-long,WU Jian-xiang, XIONG Ru-yi,ZHOU Yi-jun,ZHOU Xue-ping. Prokaryotic Expression of Coat Protein Gene S10 of Rice BlackStreaked Dwarf Virus, and Preparation and Application of Its Polyclonal Antibody [J]. , 2010, 24(1): 25-30 .
[4] XIAO Yong,LI Shuang-cheng,CHU Ming-guang,ZHOU Peng,GUAN Peng,LIU Li,WANG Ling-xia,ZHENG Ai-ping,LI Ping. Cloning,Expression and Characterization of G Protein β-subunit Gene in Rhizoctonia solani from Rice [J]. , 2008, 22(5): 541-544 .
[5] WANG Ren ,SHEN Wen-biao ,JIANG Ling ,LIU Ling-long ,ZHAI Hu-qu ,WAN Jian-min . Prokaryotic Expression,Purification and Characterization of a Novel Rice Seed Lipoxygenase Gene OsLOX1[J]. , 2008, 22(2): 118-124 .
[6] LI Ke ,WANG Shi-quan ,WU Fa-qiang ,LI Shuang-cheng ,DENG Qi-ming ,WANG Ling-xia ,LIANG Yue-yang ,LI Ping . Analysis on T4 Progeny of Transgenic Rice with Lysine-Rich Protein Gene (sb401) Mediated by Agrobacterium tumefaciens Transformation [J]. , 2008, 22(2): 131-136 .
Copyright © Chinese Journal of Rice Science 浙ICP备05004719号-5
Supported by: Beijing Magtech