Cloning Of The Cry Gene In Bacillus Thuringiensis Which Has Nematicide Activity

Chemical pesticides in the use of controlling agricultural harmful pests are significant, but due tothe excessive use of them, caused many environmental problems. Biological pesticide used in pest anddisease controlled in agriculture will get more and more widely, because of its high effective and lowtoxic. nowdays, the Bt preparations are the most widely used biological pesticides. Experiments showthat crystal protein of Bt has high pestical activity. So, further research will be on our rich Bt resources,and screening with more insecticidal activity of Bt bacteria, separating more bt strains whose gene cancode insecticidal protein, establishing Bt engineering bacterium, cultivate the turn Bt insect resistanceplants have far-reachingNematode is a kind of forestry harmful biological, it causes Pine wood nematode which is adestructive disease of pine trees, it through the pine ink spreaded. pine trees which once infectednematodes, will be dead shortly. For the moment, the prevention and control of the nematode is main tocontrol its media insects Monochamus alternatus Hope, and the most way of control is chemicalcontrol, the way of directly to control nematodes have a little difficulty and there is little report on thisboth in the domestic and abroad research. our laboratory has separated80strains of bt fromenvironment, preliminary eight strains were screened to nematicidal activities, extracting the cryprotein of the eight strains and testing their nematicidal activities, finally we selected a strain whichhas high toxic named ZJFC85strain to do our further research..We have known that the reason of ZJFC85has nematicidal activities is its cry gene can code cryproteins. In order to provide heritage materials to cultivate pine wood nematode disease resistance of thepine tree, the lab will be cloned ZJFC85strains cry gene whicn can produce cry spores. Through thereview of the literature, we designed conservative regions primers of cry gene, plasmid DNA ofZJFC85strains as PCR template, through out PCR technology,we amplifed out a conservativeregions of cry1sequence, cloned the cry1gene and send the cloning product to sequence, according tothe result of the sequence,designed cry1gene specificity primer, PCR amplificated cry1genes toget the whole length of the gene, cloned the whole length of the cry1gene, finally we got theheritage materials which is needed.