Cloning Of The Anti-alternatus Gene (cry3aa Homolog), Transformation Of Poplar Transformation

This thesis studied on Bacillus thuringiensis strain Bt886 which was separated and selected by researchers of our laboratory. According to the observation of crystal shape and the bioassay of Motschulsky and Fairmaire, Bacillus thuringiensis strain Bt886 was identified as cry3 type, and the conclusion was assured by the further study on molecular level.The 1956 base pairs full lengrh homological cry3Aa gene which was toxic to Motschulsky was cloned and sequenced. It is highly homologized to cry3Aal gene. Further sequence analysis show that only 6 base pairs of nucleotide and 2 amino acids are different between them.The homological cry3Aa gene was expressed in Escherichia coli.. And the expressed products which contain a fused peptide of 66-97 kilo-Dalton was observed by means of SDS-PAGE. Bioassay results show that the expressed protein in Escherichia coli. is toxic to Tenebrio molitor L..The homological cry3Aa gene was modified according to the characters of poplus genes and the other features of eukaryotic gene, and was artificially synthesized. The original and modified homological c/y3Aa gene were cloned into binary vector pB121 and pBin438 respectively, and transplanted into 84K clone (P. tomentos X P. glandulossa) to breed new anti-insects tree species.