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Acta Agron Sin ›› 2009, Vol. 35 ›› Issue (7): 1202-1208.doi: 10.3724/SP.J.1006.2009.01202

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

Expression of ARC1 in Vitro and Test of Interaction between ARC1 and SRK from Brassica oleracea L. in Signal Transduction Pathway of Self-Incompatibility

NIU Yi1,WANG Zhi-Min1,GAO Qi-Guo1,SONG Ming1,WANG Xiao-Jia1*,ZHU Li-Quan2*   

  1. 1Key Laboratory in Olericulture of Chongqing，Southwest University，2Plant Physiology and Biochemistry Laboratory of Southwest University，Chongqing 400716，China
  • Received:2008-12-05 Revised:2009-03-16 Online:2009-07-12 Published:2009-05-18
  • Contact: WANG Xiao-Jia,E-mail:wxj@swu.edu.cn;ZHU Li-Quan,E-mail:zhuliquan@swu.edu.cn;Tel:023-68250794;Fax: 023-68251914

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Abstract:

Self-incompatibility (SI) is a widespread mechanism in flowering plants that prevents inbreeding and promotes outcrossing. Self-pollen recognition relies on the products of genes located at the S (self-incompatibility) locus. Significant progress has been made in understanding molecular interactions allowing stigmatic cells to recognize and reject self-pollen in Brassica during the past decades. Thus, the male and female determinants responsible for the self-incompatibility (SI) response have been identified. The structural features of these molecules strongly suggest that SI response is triggered by a ligand-receptor interaction. ARC1 is anArm Repeat Containing and also a downstream gene of SRK. ARC1 interacts with SRK probably in signal transduction pathway of self-incompatibility. In this study, with an aim to testify the interaction, the coding sequences of ARC1 were amplified from stigma mRNA of Brassica oleracea L., and inserted into the expression vector pET43.1a to construct the recombinant plasma pET43.1a-ARC1, transformed E. coli(BL21) and detected expression of the recombinant protein via SDS-PAGE. Using Co-Immunoprecipitation theory and characteristic of 6×His Tag combined with Ni+ in pET43.1a-ARC1, a new method was put forward for testing the interaction between proteins. Through the method the interaction between ARC1 and SRKwas analyzed, showing that ARC1 and SRK could act with each other to combine and form a complex. This research provides a theoretical and technical bases for further analyzing the mechanism of interaction between ARC1 and SRK, for probing into interaction between ARC1 and downstream targets.

Key words: Brassica oleracea L, Signal transduction, S-locus receptor Kinase（SRK）, Arm repeat containing1（ARC1）

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