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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (04): 579-586.doi: 10.3724/SP.J.1006.2011.00579

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

Detection of Interactions between SCR and SRK in Brassica oleracea L. by Yeast Two-Hybrid System

LUO Bing1,3,XUE Li-Yan1,**,ZHU Li-Quan1,*,ZHANG He-Cui1,PENG Yi-Bo1,CHEN Song1,YANG Hong1,YANG Kun1,LI Cheng-Qiong2,WANG Xiao-Jia2,*   

  1. 1 Plant Physiology and Biochemistry Laboratory of Southwest University, Chongqing 400716, China; 2 Key Laboratory in Olericulture of Chongqing, Southwest University, Chongqing 400716, China; 3 College of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, China
  • Received:2010-09-17 Revised:2011-01-06 Online:2011-04-12 Published:2011-02-24
  • Contact: 朱利泉, E-mail: zhuliquan@swu.edu.cn, Tel: 023-68250794, Fax:023-68251914; 王小佳, E-mail: wxj@swu.edu.cn

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Abstract: For further study on mechanism of the mutual recognition between S-locus receptor kinase (SRK) and S-locus Cysteine-rich protein (SCR) in Brassica, the extracellular domains of SRK (named as eSRK) and SCR were amplified by nested PCR using Brassica oleracea L. ‘D3’ with the trait of typical self-incompatibility (SI), and the interactions between eSRK and SCR was detected by the yeast two-hybrid system. Then the full-length eSRK and the SCR were fused to the Gal4 DNA activation domain (designated pGADT7eSRK) and the Gal4 DNA binding domain (designated pGBKT7SCR) by the gene homologous recombination technique respectively. Then the pGADT7eSRK plasmid and the pGBKT7SCR were transformed into the Y187 yeast strain and the Y2HGold yeast strain respectively. The two transformed yeast strains did not exhibit autoactivation. The diploids, which were fused by the transformed Y2HGold yeast strain and the transformed Y187 yeast strain, grew on selective agar plates (SD/–Ade/–His/– Leu/–Trp/X-a-Gal/AbA), and the resulting colonies were blue, which strongly indicated that eSRK and SCR could combine with each other.

Key words: Brassica oleracea L., Self-incompatibility, S-locus receptor kinase (SRK), S-locus cysteine-rich protein (SCR), Yeast two-hybrid system

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