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《Journal of South China Agricultural University》 1992-04
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TRANSFER OF SYNTHETIC ANTIBACTERIAL PEPTIDE D GENE OF ANTHERAEA PERNYI INTO TOBACCO PLANT

Huang ZiRan Xu Fei Zhuang ChuZiong Guan ZhiWen(South China Agricultural University)Li NaiJian Xuan SiQing Que ZhiLin Dai Mian(Guangdong Academy of Agriculture Science)  
Some inducible antibacterial peptide (AP) were developed from the haemolymph of Chinese oak silkworm,Aniheraea pernyi pupae.The AP had broad-spectrum inhibition on bacteria such as wilt disease,Pseudomonas solanacearum of tobacco etc.The AP-D has been isolated and its amino acid sequence has been determined (36 peptides).We designed the nucleotides sequence of AP-D gene (122 bp) and synthesized by by DNA synthesizer.The M13 mp8 DNA was ligted with AP-D gene and transformed into E.soft JM103.The recombinant plasmid cut with Sai 1 and co24 plasmid containing CaMV 35S promotor cut with Xho I were ligated and transformed into Agrobacterium tuonefaciens by triple mating method.A Selected medium containing Km,Cm,Spc and Str was used to select the colonies which bad insert with AP-D gene.A.turnefaciens containing AP-D gene was used to infect the leaf dics of tobacco.After 4 weeks,we found some plantlets differentiation at the redges of leaf dics.These plants were resisted to Km,the nopaline detecton and Southern blot hybridization with AP-D gene probe showed that AP-D gene had been integrated into tobacco plant.The expression product should detected with its monoclonal antibody.We are now studying this in order to introduce the AP-D gene into to bacco plant.
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