A passage through in vitro culture leads to efficient production of marker-free transgenic plants in Brassica juncea using the Cre-loxP system.
The Cre-loxP site-specific recombination system was deployed for removal of marker genes from Brassica juncea (Indian mustard). Excision frequencies, monitored by removal of nptII or gfp genes in F(1) plants of crosses between LOX and CRE lines, were high in quiescent, differentiated somatic tissues but extremely poor in the meristematic regions (and consequently the germinal cells) thus preventing identification and selection of marker-free transgenic events which are devoid of both the marker gene as well as the cre gene, in F(2) progeny. We show that a passage through in vitro culture of F(1 )leaf explants allows efficient development of marker-free transgenics in the F(2) generation addressing current limitations associated with efficient use of the Cre/loxP technology for marker gene removal.
Arumugam
N
N
Centre for Genetic Manipulation of Crop Plants, University of Delhi South Campus, Benito Juarez Road, New Delhi 110 021, India.
Gupta
Vibha
V
Jagannath
Arun
A
Mukhopadhyay
Arundhati
A
Pradhan
Akshay K
AK
Burma
Pradeep Kumar
PK
Pental
Deepak
D
D001937:Brassica / Q000201:enzymology / Q000235:genetics*; D003433:Crosses, Genetic; D005818:Genetic Engineering; D005819:Genetic Markers; D019426:Integrases / Q000235:genetics*; D030821:Plants, Genetically Modified; D046509:Tissue Culture Techniques
D005819:Genetic Markers; C045073:Cre recombinase; D019426:Integrases