巴西橡胶树HbNAC1转录因子互作顺式作用元件的筛选

Screening of Cis-elements Interacted with HbNAC1 of Hevea brasiliensis

巴西橡胶树HbNAC1转录因子可能与脱水信号诱导的橡胶乳管分化相关,此外,HbNAC1能结合Hb SRPP的启动子并调控该基因的表达,因此可能是一个橡胶产排胶调控相关基因。笔者首先构建多个顺式作用元件的p Ab Ai酵母单杂载体,然后构建HbNAC1转录因子的p GADT7-Rec2-HbNAC1载体,2种载体共转化Y1H Gold酵母菌株,然后通过酵母单杂交筛选与HbNAC1互作的顺式作用元件。结果表明,HbNAC1可结合JRE,CACG和ABRE顺式作元件并激活报告基因AUR1-C的表达。本研究结果为鉴定HbNAC1转录因子的靶基因,研究其下游基因的表达调控提供重要的线索。

HbNAC1 has been identified as a key transcriptional factor related with laticifer differentiation induced by local dehydration in Hevea brasiliensis. Moreover,HbNAC1 could bind the promoter of and regulate the gene expression of Hb SRPP,suggesting that HbNAC1 might be involved in latex biosynthesis of Hevea brasiliensis. Several cis-elements were inserted into p Ab Ai vector of Yeast-one-hybrid（ Y1H）,and a vector p GADT7-Rec2-HbNAC1 was then constructed for HbNAC1. Both vectors were co-transferred into Y1 H Gold yeast strain to select cis-elements interacted with HbNAC1. Y1 H hybridization results showed that HbNAC1 were bound to ciselements JRE,CACG and ABRE and activated the expression of AUR1-C report gene. These results are very important for identification and characterization of target genes in downstream in the future.