A study by Chawla and colleagues documented how site-specific integration in rice exhibited stable gene expression over multiple generations. Concerns over the presence of antibiotic resistance genes in the food supply and their escape into the environment can be relieved through the use of recombinase technology to excise unwanted DNA from the genome of genetically engineered (GE) crops prior to marketing or release. Recombinase-mediated genetic engineering provides a favorable direction for enhancing the precision of biotechnological approaches. Publicly acceptable forms of biotechnology offer an avenue for meeting these demands. Plant biotechnology has a role in addressing global needs for food, fiber and fuel, by developing new crop varieties with increased pest resistance, biofortification, and abiotic stress tolerance. The precise site-specific deletion by phiC31 in planta demonstrates that the recombinase can be used to remove selectable markers or other introduced transgenes that are no longer desired and therefore can be a useful tool for genome engineering in plants. The phiC31 system performs site-specific recombination in germinal tissue, a prerequisite for generating stable lines with unwanted DNA removed.
#MACVECTOR ATTB RECOMBINATION MANUAL#
We further verified that the genomic excision was conservative and that introduction of a functional recombinase can be achieved through secondary transformation as well as manual crossing. The phiC31 recombinase excised the attB and attP-flanked DNA, and the excision event was detected in subsequent generations in the absence of the phiC31 gene, indicating germinal transmission was possible. In this work, the phiC31 recombinase gene was placed under the control of the Arabidopsis OXS3 promoter and introduced into Arabidopsis harboring a chromosomally integrated attB and attP-flanked target sequence. Previously, we characterized the phiC31 catalytic activity and modes of action in the fission yeast Schizosaccharomyces pombe. The large serine recombinase phiC31 from broad host range Streptomyces temperate phage, catalyzes the site-specific recombination of two recognition sites that differ in sequence, typically known as attachment sites attB and attP.
0 kommentar(er)
