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Acta Agron Sin ›› 2014, Vol. 40 ›› Issue (07): 1174-1181.doi: 10.3724/SP.J.1006.2014.01174

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Characterization of Brassica juncea Zinc Finger Protein Transcription Factor Gene Bj26

JIA Shuang-Wei1**,GAO Ying1*,ZHAO Kai-Jun1,*   

  1. National Key Facility for Crop Gene Resources and Genetic Improvement / Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2014-01-27 Revised:2014-04-16 Online:2014-07-12 Published:2014-05-16
  • Contact: 赵开军, E-mail: zhaokaijun@caas.cn, Tel:010-82105852

Abstract:

 

Zinc-finger proteins, forming an important transcriptional factor family, are involved in gene transcriptional regulation, development and stress-responses in plants. The previous studies revealed that BjC-P is a fungus-inducible promoter from Brassica juncea and W-box-like-4 is the core element responsive to fungal infection. Here, we report the molecular cloning and characterization of a zinc finger protein-encoding gene (designated Bj26). Bj26 was screened out from the Brassica juncea cDNA library by the Yeast One-Hybrid System. Bioinformatic analysis showed that Bj26 contains open reading frame of 735 bp that encodes a new C2H2-type zinc finger protein with an isoelectric point (pI) of 9.2 and a molecular weight of 26.6 kD. The protein consists of two typical zinc-finger domains and contains two conserved QALGGH amino acid sequences. Subcellular localization showed that Bj26 is located in the nuclear. The histochemical and quantitative GUS assays, through transient gene expression in Nicotiana benthamiana leaf, showed that Bj26 can bind to the core element W-box-like-4 and activate the function of BjC-P. qRT-PCR analysis showed that the gene Bj26 expression obviously increased under the induction of the fungal elicitor (Hexa-N-Acetyl-Chitohexaose). CDS alignments and phylogenetic analysis of Bj26 and other C2H2-type proteins from Arabidopsis thaliana (At) and Oryza sativa (Os) (MEGA 6) showed Bj26 shares high similarity with that from Arabidopsis thaliana (At). All of the above results suggested that Bj26 protein mediates the process of plant response to fungal pathogen.

Key words: Yeast One-Hybrid System, C2H2-type zinc finger protein, Bj26, Fungal-induction, Transient gene expression, qRT-PCR

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