Cloning and Functional Analysis of Xylem Specific Promoter Deletion Fragment  

作　　者：Yajing LUO Liang WANG Kejiu DU Shuang ZHANG 

机构地区：[1]Hebei Agriculture University,Baoding 071000,China [2]Baoding Jingxiu Park Management Office,Baoding 071051,China [3]Hebei Key Laboratory of Forest Tree Gennplasm Resources and Forest Protection,Baoding 071000,China

出　　处：《Agricultural Biotechnology》2016年第6期1-3,9,共4页农业生物技术（英文版）

基　　金：Supported by National Science Foundation of China(31000305);Fund from Education Department of Hebei Province(QN2015182);Innovation Fund for Forestry Discipline of Hebei Agricultural University(LXXK2014-1);Fund for Overseas Research and Study by Young and Middle-aged Backbone Teachers in Hebei Agriculture University(JWYX2015)

摘　　要：Xylem-specific promoter could regulate efficient expression of foreign genes in xylem. Deletion analysis method was applied to obtain 5＇ deletion promoter fragments MU2 andMU4 with different distances from transcription start site of xylem-specific promoter MDCcsAP through PCR procedure. Then CaMV35S promoter in plant expression vector pBI121 was replaced by amplified fragments. The recombinant plasmids fused with corresponding gene segments and GUS reporter gene were obtained and transformed into Agrobacterium. The results of the tobacco transient transformation test showed that MU2 andMU4 had the promoter function, and both of their activity were significantly higher than that of CaMV35S promoter, and they had the characteristics of xylem tissue specificity as well. Research on xy- lem-specific promoter structure and function could lay a foundation for the determination of necessary components to the xylem tissue specificity and cis-acting ele- ments with induction activity, so as to enable the regulation of efficient expression of exogenous genes in specific time and specific tissues in plants through these cis-acting elements.Xylem-specific promoter could regulate efficient expression of foreign genes in xylem. Deletion analysis method was applied to obtain 5＇ deletion promoter fragments MU2 andMU4 with different distances from transcription start site of xylem-specific promoter MDCcsAP through PCR procedure. Then CaMV35S promoter in plant expression vector pBI121 was replaced by amplified fragments. The recombinant plasmids fused with corresponding gene segments and GUS reporter gene were obtained and transformed into Agrobacterium. The results of the tobacco transient transformation test showed that MU2 andMU4 had the promoter function, and both of their activity were significantly higher than that of CaMV35S promoter, and they had the characteristics of xylem tissue specificity as well. Research on xy- lem-specific promoter structure and function could lay a foundation for the determination of necessary components to the xylem tissue specificity and cis-acting ele- ments with induction activity, so as to enable the regulation of efficient expression of exogenous genes in specific time and specific tissues in plants through these cis-acting elements.

关 键 词：PROMOTER Deletion analysis Vector construction GUS activity analysis 

分 类 号：Q943.2[生物学—植物学]