Of Tat-cry1ac, The Fusion Protein Screening Positive In Tobacco, Arabidopsis Plants To Diamondback Moth Control Study

DBM(Plutella xylostella)belongs to Lepidoptera moth families,are mainly pests of cruciferous vegetables,they are distributed more than 80 countries in the world.It becomes a great threat of brassicaceous vegetable production in China.It is reported that DBM produces high resistance to Bt insecticidal gene in multiple countries or regions.Protein transduction domain(trans-activator of transcription, TAT)is a kind of auto matically penetrate the cell membrane, and can serve as vectors of other biological macromolecules into the cell membrane to bring the section with in the arginine-rich peptides.A section of synthetic TAT gene is used in this paper and with laboratory-preserved cry1Ac insecticidal gene as a basis, pBI121-Cry1Ac,pBI121-TAT-Cry1Ac pBI121-Cry1Ac-YFP,pBI121-TAT-Cry1Ac-YFP, four kinds of recombinant plasmids are constructed by yellow fluorescent protein (YFP) as a marked gene.Completion of the four fusion mediated gene vector in model plant tobacco, Arabidopsis to explore genetic transformation conditions, transgenic tobacco and Arabidopsis plants positive clones are successfully screened by PCR,RT-PCR molecular detection of Cry1Ac gene specific primers genomes and Western-blot analysis of CrylAc toxin protein;positive plant protein is extracted,then completed quantitative expressing protein of fusion gene in plants by ELISA.Resistance and sensitive DBM are used in experiments as sources, positive tobacco plant protein is extracted,and is bioassaying in room by positive young leaves of arabidopsis plants. The results show that the TAT protein transduction domain have significantly the role of synergy and grams of the Cry1Ac, the positive expression of the tobacco plant efficiency Plutella sensitive fusion protein range:3.260～3.296, confrontational moth efficiency range:2.107～2.169; Arabidopsis plants positive for expression of fusion proteins on the sensitive moth efficiency interval:3.260～3.296, efficiency relative to the diamondback moth resistance range:2.107～2.169.