拟南芥COI 1互作基因的分离

为进一步阐明茉莉素调控植物生长发育的分子机理，为作物抗性和雄性不育基因工程提供新思路，以COI1为诱饵，利用酵母双杂交分析体系筛选拟南芥cDNA文库，得到300多个阳性克隆，经PCR，RsaⅠ酶切PCR产物以后归为12群，从代表菌落提取pB42AD cDNA重组质粒进行了测序，完成了序列分析，得到ASK1，COAP2，COAP3，COAP4等12个基因，其中ASK1，COAP2，COAP3等基因与Skp1等重要基因有较高的同源性。

Isolation of Arabidopsis Genes Interacting with COI 1

The authors used COI 1 as bait to screen Arabidopsis cDNA library in yeast two-hybrid system. More than 300 putative positive colonies have been isolated,which belonged to 12 distinct groups. Representative cDNAs from each group were sequenced. DNA database searching showed that one of these genes, represented by 59 positive colony, was identical to the previously identified Arabidopsis Skp1 like protein ASK1 (formerly named as AtSkp 1).Other putative COI 1 associated proteins were designated as COAP2, 3 to 12, respectively. COI 1 interacted strongly with ASK1, COAP2 and COAP3.Presence of the F-box motif in COI 1 led us to the prediction that COI 1 might interact with Skp 1-like protein to form SCF ubiquitin complex.Indeed,ASK1,COAP2 and COAP3 identified in the yeast two-hybrid screen are similar to the Skp 1 protein from yeast and human.The authors therefore focused their analysis on ASK1,COAP2, COAP3 and COAP4 with the complementary biochemical and molecular approaches.