Marker gene excision in transgenic Brassica napus via Agrobacterium-mediated Cre/lox transient expression system

作     者：Pandi Wang Xiaojuan Xiong Ping Fu Gang Wu Fang Liu Pandi Wang;Xiaojuan Xiong;Ping Fu;Gang Wu;Fang Liu

作者机构：Key Laboratory of Biology and Geneics Improvement of Oil CropsMinistry of Agriculture and Rural AffairsOil Crops Research InstituteChinese Academy of Agricultural SciencesWuhanChina 

出 版 物：《Oil Crop Science》 (中国油料作物学报(英文版）)

年 卷 期：2020年第5卷第2期

页      面：96-103页

学科分类：09[农学] 0901[农学-作物学] 

基　　金：supported by the National Program of Transgenic Variety Development of China(2018ZX08010-05B and 2019ZD080018) the Central Public-interest Scientific Institution Basal Research Fund Major Research Project of CAAS Science 

主　　题：Brassica napus Marker-free Cre/lox Cross Auto-excision Herbicide resistance 

摘      要：Oilseed rape(Brassica napus L.)is one of the most important oil crops in the ***,study on marker-free transgene of *** for bio-safety purpose is limited in this allotetraploid *** order to advance marker gene excision research,a newly designed Cre/lox system combining crossing and auto-excision strategy was introduced into *** system consists of 2 sets of independent vectors including pC35 Spro::T7 RP carrying T7 RNA polymerase and pCT7 pro::Cre carrying T7 promoter *** hybridization of 2 according types of transgenic ***,marker gene would be removed as T7 RNA polymerase facilitate T7 promoter to promote Cre gene *** 52 and 46 positive To transgenic lines of these 2 vectors were obtained after identification by PCR and test trip.T1 plants from 3 T0 positive pC35 Spro::T7 RP lines and 2 T0 positive pC35 Spro::T7 RP lines were identified as single copy according to segregation ratio and were chosen for ***,expression of CP4 EPSPS(glyphosate resistance gene)and OXY(bromoxynil resistance gene)were not found in F1 progeny,which proved that the excision was not *** possible reasons for our limited success were investigated and detailed analyses were *** this system is not applicable for generating transgenic *** free from selectable marker gene,it provided valuable experience and clue for further improvement of this *** other advantages and further improvement will be progressed in future work.