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Acta Agron Sin ›› 2008, Vol. 34 ›› Issue (06): 934-943.doi: 10.3724/SP.J.1006.2008.00934


Test of Interaction between THL1 and SRK from Brassica oleracea L. in Self-Incompatibility Signaling Process

GAO Qi-Guo1,SONG Ming1,NIU Yi1,YANG Kun2,ZHU Li-Quan2*,WANG Xiao-Jia1*   

  1. 1 Key Laboratory in Olericulture of Chongqing, Southwest University, Chongqing 400716; 2 Plant Physiology and Biochemistry Laboratory of Southwest University, Chongqing 400716, China
  • Received:2007-11-14 Revised:1900-01-01 Online:2008-06-12 Published:2008-06-12
  • Contact: ZHU Li-Quan

Abstract: The SRK and THL1 from Brassica oleracea L. act probably with each other in self-incompatibility signal process. In present study, with an aim to testify the interaction, the coding sequences of THL1 and SRK kinase domain(named as SRKE1) were amplified from stigma mRNA of Brassica oleracea L., and inserted into the expression vector pET43.1 and yeast expression vec-tor pPIC9K to construct the recombinant plasma pET43.1-THL1, pET43.1-SRKE1, and pPIC9K-SRKE1. The recombinant protein were expressed in E. coli(BL21) and Pichia pastoris (GS115), respectively. Thioredoxin activity was tested by the ability of THL1 reducing insulin, showing that THL1 had oxidation/reduction activity. A new method was put forward for testing the interaction between proteins, through the method the interaction between SRKE1 and THL1 was analyzed, showing that SRKE1 and THL1 could act with each other to combine and form a stable complex. This research provides a theoretical and technical bases for fur-ther analysis of the interaction mechanism between SRK and THL1.

Key words: S-locus receptor kinase, Thioredoxin-like protein, Pichia pastoris

[1] LUO Bing, XUE Li-Yan, SHU Li-Quan, ZHANG He-Cui, PENG Yi-Bei, CHEN Song, YANG Hong, YANG Hun, LI Cheng-Qiong, WANG Xiao-Jia. Detection of Interactions between SCR and SRK in Brassica oleracea L. by Yeast Two-Hybrid System [J]. Acta Agron Sin, 2011, 37(04): 579-586.
[2] NIU Yi,WANG Zhi-Min,GAO Qi-Guo,SONG Ming,WANG Xiao-Jia,ZHU Li-Quan. Expression of ARC1 in Vitro and Test of Interaction between ARC1 and SRK from Brassica oleracea L. in Signal Transduction Pathway of Self-Incompatibility [J]. Acta Agron Sin, 2009, 35(7): 1202-1208.
[3] LIU Dong; ZHU Li-Quan; WANG Xiao-Jia. Cloning and Characterization of Encoding Sequence of SRK-Binding Protein ARC1 from Brassica oleracea L. in Self-incompatibility Signaling Process [J]. Acta Agron Sin, 2004, 30(05): 427-431.
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