作物学报2012，Vol.38Issue(7)：1328-1333,6.DOI:10.3724/SP.J.1006.2012.01328
芥菜AP1基因体外表达及其与FLC相互作用的验证
Expression of Floral Meristem Identity Gene AP1 in vitro and Validation of Interaction between AP1 and FLC in Brassicajuncea Coss.(Mustard)
摘要
Abstract
There exists a possible direct interaction between floral meristem factor API and flowering pathway central regulator FLC in Brassica juncea Coss. To further prove the interactive mechanism between API and FLC, the protein interaction in vitro was testified in Brassica juncea Coss. (Mustard). The cDNA of API gene isolated using homologous cloning techniques from mustard "QJ" was 790 bp, encoding 256 amino acids. API 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 (P), 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, recombi-nant plasmid pET43.1a-API was constructed, transformed to E. Coli (BL21) and then induced protein expression by IPTG With the characteristics of 6xHis tag in fusion protein of pET43. 1a-API which could combine with Ni+, the interaction between API and FLC was analyzed via SDS-PAGE. The results showed that API 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 Brassicajuncea.关键词
芥菜/AP1基因/FLC/蛋白互作Key words
Brassicajuncea Coss./ API gene/ FLC/ Protein interaction引用本文复制引用
汤青林,许俊强,宋明,王志敏..芥菜AP1基因体外表达及其与FLC相互作用的验证[J].作物学报,2012,38(7):1328-1333,6.基金项目
本研究由国家自然科学基金项目(31000908),中央高校基本科研业务费专项资金(XDJK2009C124,XDJK20128020),重庆市自然科学基金(2009BB1307,2011BA1002)和国家重点基础研究发展计划(973计划)项目(2012CB1139000)资助. （31000908）
