<FONT face="Verdana">Cloning， Expression  and Sequence Analysis of G Protein  β Subunit Gene of Rice  False Smut Pathogen <EM>Ustilaginoidea virens</EM></FONT>

LIU  Lian-meng,WANG  Ling,HUANG  Wen-wen,LIU  En-yong,  HUANG  Shi-wen*

doi:10.3969/j.issn.1001-7216.2010.04.04

2010 , Vol. 24 >Issue 4: 353 - 359

DOI: https://doi.org/10.3969/j.issn.1001-7216.2010.04.04

研究报告

Cloning， Expression and Sequence Analysis of G Protein β Subunit Gene of Rice False Smut Pathogen Ustilaginoidea virens

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China National Rice Research Institute， Hangzhou 310006, China； *Corresponding author, E-mail: swhuang666@sohu.com

Received date: 1900-01-01

Revised date: 1900-01-01

Online published: 2010-07-10

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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.

Key words： Ustilaginoidea virens; heterotrimeric G protein; G protein beta subunit gene; thermal asymmetric interlaced PCR; prokaryotic expression

Cite this article

LIU Lian-meng,WANG Ling,HUANG Wen-wen,LIU En-yong, HUANG Shi-wen* . Cloning， Expression and Sequence Analysis of G Protein β Subunit Gene of Rice False Smut Pathogen Ustilaginoidea virens[J]. Chinese Journal OF Rice Science, 2010 , 24(4) : 353 -359 . DOI: 10.3969/j.issn.1001-7216.2010.04.04

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