| Pests seriously affect agricultural production, crop yields and quality,restrict agricultural economic stability and development.It has long been widely used chemical insecticides to control pests.But as time goes on, the shortcomings of chemical pesticides gradually exposed.The use of plants insect-resistant genes project cultivation of crops can avoid pest damage,also has the advantage of lower cost,specificity etc,thus attracting much attention.The plants insect-resistant genes project becomes the focus of agricultural biotechnology research.Genetic engineering of plant resistance to insects is one of the most important research of plant genetic engineering.At present,there are many insect-resistant genes have been isolated from bacteria,plants and insects itself.Many insect-resistant transgenic plant are in the progress of field trials,and some variety is commercialization.The production of these transgenic plant showed a great promise.However,there are also some potential problems in the application of transgenic plants,it is necessary to develop some new,highly efficient insect-resistant genes to solve these problems.This research is carried out with this background.The main results are:(1):Use the castor genome as a template,ricin toxin gene was amplified.Then constructed the cloning vector,prokaryotic expression vector.After the induction of IPTG, E.coli BL21 which has been transformed into the recombinant plasmid expressed the objective protein.SDS-PAGE showed that the target protein expressed as inclusion bodies.(2): We cloned and expressed the ricin B-chain,and through the western blot,purification and other methods to identify this protein,and laid the foundation for further validate this protein function.(3):The plant expression vector P-En was constructed. The DNA fragment in the coding region of TnGV-En gene was in downstream of CaMV35S promoter of vector pBI121, restriction enzyme cutting confirmed that the gene had been correctly inserted in the vector. |
PDF Full Text Request