Analysis of effectiveness of the Cre/loxp site-specific recombinase system through subsequent generations of Arabidopsis thaliana transformants
Journal Title: Вісник Українського товариства генетиків і селекціонерів - Year 2015, Vol 13, Issue 1
Abstract
Aim. The aim of the work was to analyze the effectiveness of the Cre/loxP site-specific recombinase system through subsequent generations of Arabidopsis thaliana transformants. Methods. The increase in the number of marker-free Arabidopsis thaliana transformants was studied through several subsequent generations. Genetic transformation of plants was carried out using the Cre/loxP site-specific recombinase system under the control of 35S promoter. For this purpose two types of DNA constructs were designed that contain the marker gene gus and the recombinase gene cre flanked by loxP sites. Hygromycin resistance gene hptII in both constructs was moved out of the loxP sites, and in the case of excision event it remained in plant genome. The selection of transformants was performed on Murashige and Scoog medium with hygromycine (100 mg/l). The recombinase-mediated excision event of marker genes in genome was determined by histochemical analysis and molecular genetic analysis of three subsequent generations of transformants. Results. The number of marker-free transformants was increased with each subsequent generation for both types of DNA-constructs. Moreover, the reduction in the total amount of integrated T-DNA per genome to a minimum was identified by molecular-genetic analysis that indicates the ability of the Cre/loxP site-specific recombinase system to control the number of the integration events in plant genome. Conclusions. The developed approach is simple in usage and effective tool for obtaining marker-free transformed plants.Keуwords: selective marker genes, removal of marker genes, site-specific recombinase.
Authors and Affiliations
A. S. Sekan
Keywords
Related Articles
- EP ID EP613850
- DOI -
- Views 80
- Downloads 0
Vitaly Arnoldovych Kordium (to 85th anniversary)
Професор Ю.П. Мірюта – педагог і вчений (до 110-річчя від дня народження)
Висвітлено, поряд з відомими, нові біографічні та архівні матеріали про навчання, педагогічну та наукову діяльність видатного українського генетика із вавиловської плеяди вчених професора Юрія Петровича Мірюти, незалежно...
New interactions of invadopodia scaffold protein TKS5 with proteins that take part in actin cytoskeleton reorganization, endo-/exocytosis and membrane remodeling
Aim. TKS5 is a key scaffold protein of invadopodia. In its absence, the cells completely lose the ability to form invadopodia. This fact makes TKS5 a potential target for cancer cure and one of the central proteins in th...
A few words on the occasion of the three anniversaries of Ukrainian genetic science
ПЦР-анализ генов фотопериодической чувствительности у сортов мягкой озимой пшеницы селекции Белоцерковской опытно-селекционной станции
Цель. Определение аллельного состояния генов чувствительности к фотопериоду Ppd-1 у 16 сортов мягкой озимой пшеницы селекции Белоцерковской опытно-селекционной станции и детекция гаплотипов по гену Ppd-D1 согласно принят...