缺铁应答基因FOX1上游调控基因的筛选  

Identification of promoter binding proteins of FOX1 gene using yeast one-hybrid system in Chlamydomonas reinhardtii

作　　者：李亚军[1] 费小雯[2] 邓晓东[1] Yajun Li;Xiaowen Fei;Xiaodong Deng(Institute of Tropical Bioscience and Biotechnology of Chinese Academy of Tropical Agricultural Sciences ＆ Key Laboratory of Biology and Genetic Resources of Torpical Crops, Ministry of Agriculture, Haikou 571101, China;Department of Biochemistry, Halnan Medical College, Halkou 571199, China)

机构地区：[1]中国热带农业科学院热带生物技术研究所农业部热带作物生物学与遗传资源利用重点实验室,海南海口571101 [2]海南医学院理学院,海南海口571199

出　　处：《生物技术》2018年第2期103-107,共5页Biotechnology

基　　金：国家自然科学基金项目("莱茵衣藻缺铁应答基因Femu2p功能解析";No.31160050;"莱茵衣藻MAPKKK激酶Femu9p介导的缺铁信号转导途径研究";No.31360051)

摘　　要：[目的]筛选与莱茵衣藻FOX1基因启动子结合的调控基因序列。[方法]利用酵母单杂交的方法,构建pHIS2.1-FOX1诱饵载体,并将其转入到酵母菌株Y187中,在SD/-Trp/-His/-Leu/50 mmol/L 3-AT筛选培养基上挑选酵母阳性转化子,PCR克隆阳性转化子的序列并测序,利用Blastx程序检索phytozome莱茵衣藻基因组数据库,获得阳性转化子序列的同源基因信息。[结果]18个酵母阳性转化子测序成功,其同源基因主要有CTR型铜离子转运体(CTR2)、核糖体蛋白,和参与光合作用、氧化还原反应和物质运输等方面的功能蛋白。[结论]利用酵母单杂交技术筛选到缺铁应答基因FOX1潜在的上游调控基因。[Objective]Screening the binding proteins of FOX1 gene in Chlamydomonas reinhardtii.[Methods]The bait vector pHIS2.1-FOX1 was constructed and transfomed into yeast strain Y187.The positive yeast clones were selected on SD/-Trp/-His/-Leu/50 mmol/L 3-AT media with the MatchmakerTMone-hybrid library construction screening kit.The sequences of putative binding proteins of FOX1 gene were obtained by using PCR and sequencing.The sequences were blasted in Phytozome Chlamydomonas reinhardtii database.[Results]Total 18 yeast positive clones were successfully sequenced,and the genes encoding proteins were homologous to CTR type copper ion transporter（CTR2）,ribosomal proteins and several functional proteins related to photosynthesis,redox reactions and transporters.[Conclusion]The putative upstream genes of FOX1 were identified by using yeast one-hybrid method.

关 键 词：酵母单杂交 莱茵衣藻 缺铁应答 FOX1基因 启动子 上游调控基因 

分 类 号：Q781[生物学—分子生物学] Q946.2