公开(公告)号：US5827684A

公开(公告)日：1998-10-27

申请号：US231342

申请日：1994-04-19

申请人： Kotikanyadanam Sreekrishna ,  William D. Prevatt ,  Gregory P. Thill ,  Geneva R. Davis ,  Patricia Koutz ,  Kathryn A. Barr ,  Sharon A. Hopkins

发明人： Kotikanyadanam Sreekrishna ,  William D. Prevatt ,  Gregory P. Thill ,  Geneva R. Davis ,  Patricia Koutz ,  Kathryn A. Barr ,  Sharon A. Hopkins

IPC分类号： A01N63/00 ,  A01N63/02 ,  A01N63/04 ,  C07K14/32 ,  C12N1/19 ,  C12N15/09 ,  C12N15/81 ,  C12P21/02 ,  C12R1/07 ,  C12R1/84 ,  C12N15/32 ,  C12N15/63 ,  C12N21/02

CPC分类号： C12N15/815 ,  A01N63/04 ,  C07K14/32

摘要： A method for producing one or more Bacillus toxin polypeptides by culturing methylotrophic yeast cells which have a gene(s) capable of expressing the Bacillus toxin polypeptide(s) in such cells under conditions that the gene(s) is/are transcribed is provided. The toxin polypeptide encoding segment of the gene(s) has a G+C content of about 40%-55%, and preferably comprises methylotrophic yeast codons. The preferred species of yeast for expressing such synthetic Bacillus toxin gene(s) is Pichia pastoris. Bacillus toxin polypeptides encoded by synthetic genes are expressed at high levels in transformed methylotrophic yeast cells. The toxin expressing cells may be administered as live cells or heat-killed whole cells to provide an insecticidal composition for killing susceptible insect larvae. Also provided by the present invention are DNAs capable of transforming methylotrophic yeast to express one or more Bacillus toxin polypeptides, cultures of such yeast cells transformed with such DNAs and novel Bacillus toxin polypeptides made by the method of the invention. The transformed yeast cells of the present invention are readily ingested as food by insect larvae which are susceptible to the toxin polypeptides.

摘要翻译： 提供了一种通过培养具有能够在基因被转录的条件下能够在这些细胞中表达芽孢杆菌毒素多肽的基因的甲基营养酵母细胞产生一种或多种芽孢杆菌毒素多肽的方法。 编码基因片段的毒素多肽的G + C含量为约40％-55％，优选包含甲基营养酵母密码子。 用于表达这种合成芽孢杆菌毒素基因的酵母的优选种类是巴斯德毕赤酵母。 由合成基因编码的芽孢杆菌毒素多肽在转化的甲基营养酵母细胞中以高水平表达。 表达毒素的细胞可以作为活细胞或热灭活的全细胞施用，以提供用于杀灭易感昆虫幼虫的杀虫组合物。 本发明还提供能够转化甲基营养酵母以表达一种或多种芽孢杆菌毒素多肽的DNA，用这种DNA转化的这种酵母细胞的培养物和通过本发明的方法制备的新的芽孢杆菌毒素多肽。 本发明的转化酵母细胞易于被毒素多肽易感的昆虫幼虫摄取为食物。