Co-expression of GUS and GFP in hairy roots of Nicotiana tabacum Wisconsin 38
Date Issued
2011
Date
2011
Author(s)
Huang, Yu
Abstract
To establish a platform of Agrobacterium rhizogenes-mediated multiple-gene transformation, we proposed and evaluated different strategies in this study. Two reporter genes are carried by two separate plasmids within different transformed A. rhizogenes (2AR) and within one transformed A. rhizogenes (2BV). Two reporter genes are also constructed in one T-DNA (1TD) and in different T-DNA (2TD) in one binary vector. In the study, we compare the hairy root inducing rate, genes co-transforming rate and the expression of reporter genes-GUS and GFP. Hairy root induction rate of the four groups were all over 90%, without difference from wild type A. rhizogenes. The transformation efficiency of 2AR, 2BV, 1TD and 2TD was 65.4%, 40%, 78.6%, and 82.1%, respectively. Our results showed that all of the four strategies can co-express GUS and GFP in tobacco hairy roots. The GUS expression of 2AR, 2BV, 1TD and 2TD ranged from 0 to 259.1, from 0.2 to 698.9, from 0.7 to 112.3, and from 0.6 to 698.1 nmol MU min-1 mg-1 protein, respectively. Meanwhile, the transgenic hairy roots expressed GFP ranging from 5.5 to 6333.9, from 0.4 to 8758.7, from 1.3 to 1923.5, and from 0.7 to 8459.3 ng mg-1 protein, respectively. 1TD and 2TD were better strategies to gain co-expression hairy roots, while all of the four strategies were able to transform multiple genes into plants.
Subjects
A. rhizogenes
tobacco hairy root
co-transformation
Type
thesis
File(s)![Thumbnail Image]()
Loading...
Name
ntu-100-R98b47114-1.pdf
Size
23.32 KB
Format
Adobe PDF
Checksum
(MD5):d7e456cd0cc35129ec7316f1445f182e