Construct Of CYP2E1and GST Genes Binaiy Expression Vector And Its Expression In The Alfafa

With the process of industrialization and further development of urbanization, Theenvironmental pollution of China is growing. The type and quantity of pollutants in theenvironment is increasing which is seriously damaging the economic development andhuman health. At present, the methods which used to remediate soil pollution includephysical/chemical remediation and bioremediation categories. Phytoremediation is theuse of green plants to remove pollutants from the environment. Compared with thetraditional repair techniques, phytoremediation technology for more rapid, efficient,low cost, and easy to promote, has shown a good development prospects in recentyears.This study transduced the two genes into alfalfa using transgenic technology toeffectively repair soil heavy metal-organic multi-pollution conditions. GlutathioneS-transferase gene GST and cytochrome oxidase gene CYP2E1respectively relatewith resistant and accumulation of heavy metals and organic compounds. At present,there is no same report about this research.In this paper, we constructed a expression vector which carring the CYP2E1gene:the Gus gene in the vector PBI121was replaced by the exogenous gene at first,construct P35S-CYP2E1-T35S expression cassettes which can expressed highly in thealfalfa. Then, we transduce the recombination plasmids to LBA4404with GST gene bythe freeze thawing method.The experiment used "Algonquin" alfalfa’s hypocotyl as transformation acceptor,through the tissue culture researching, screened the induction and differentiationconditions of embryogenic callus. Further, optimized the genetic-transformation system which suit to the "Algonquin" alfalfa. Alfalfa embryo callus were transformedwith GST gene and CYP2E1gene by Agrobacterium-mediate transformation.Inoculated callus were cultured successively on cocultive media, selective media.240Kan-resistant alfalfa plants were obtained.After molecular level detection with the approaches of PCR and RT-PCR to the240selected resistant plants, resistant functions to the heavy metals and organiccompounds of the transgenic plants were identified. The analysis showed the resistantalfalfa plants integrated GST gene and CYP2E1gene successfully and expressedeffectively.