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

Oil Crop Science 2020年第2期5卷 96-103页

作者： Pandi Wang Xiaojuan Xiong Ping Fu Gang Wu Fang Liu Key Laboratory of Biology and Geneics Improvement of Oil Crops Ministry of Agriculture and Rural AffairsOil Crops Research InstituteChinese Academy of Agricultural SciencesWuhanChina

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.

关键词： Brassica napus Marker-free Cre/lox Cross auto-excision Herbicide resistance

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