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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (07): 1328-1333.doi: 10.3724/SP.J.1006.2012.01328

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Expression of Floral Meristem Identity Gene AP1 in vitro and Validation of Interaction between AP1 and FLC in Brassica juncea Coss. (Mustard)

TANG Qing-Lin**,XU Jun-Qiang**,SONG Ming*,WANG Zhi-Min   

  1. College of Horticulture and Landscape Architecture, Southwest University / Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education / Key Laboratory of Olericulture, Chongqing 400715, China
  • Received:2011-09-30 Revised:2012-04-16 Online:2012-07-12 Published:2012-05-11
  • Contact: 宋明, E-mail: swausongm@yahoo.com.cn

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Abstract: There exists a possible direct interaction between floral meristem factor AP1 and flowering pathway central regulator FLC in Brassica juncea. Coss. To further prove the interactive mechanism between AP1 and FLC, the protein interaction in vitro was testified in Brassica juncea Coss. (mustard). The cDNA of AP1 gene isolated using homologous cloning techniques from mustard “QJ” was 790 bp, encoding 256 amino acids. AP1 belongs to MIKC type protein with MADS domain and K-box known from analysis software. The conserved MADS domain had two alpha helixes (a) and two beta-sheets (b), and one amino acid site in the first a helix was not conserved. The K-box had three alpha helixes (a), and one amino acid site in the first and second a helixes was not conserved, respectively, but four amino acid sites in the third a helix were not conserved. Furthermore, recombinant plasmid pET43.1a-AP1 was constructed, transformed to E. coli (BL21) and then induced protein expression by IPTG. With the characteristics of 6×His tag in fusion protein of pET43.1a-AP1 which could combine with Ni+, the interaction between AP1 and FLC was analyzed via SDS-PAGE. The results showed that AP1 and FLC could act with each other to combine and form a complex. This research provides theoretical and technical bases for further analyzing the interaction mechanism of AP1-FLC protein complex and the molecular regulation of floral meristem in Brassica juncea.

Key words: Brassica juncea Coss., AP1 gene, FLC, Protein interaction

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