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苏云金芽孢杆菌高效菌株cry基因分析及多价基因组合的研究
cry GENE ANALYSIS AND MULTI cry GENE COMBINATIONS OF SEVERAL HIGHLY TOXIC BACILLUS THURINGIENSIS STRAINS
【摘要】 对本室筛选的高活性B.t.野生株进行了cry 基因检测,Southern杂交证明cryIAc和cry1C位于质粒上.利用接合转移和电穿孔转化等方法对这些菌株进行了遗传改良研究.结果表明:高效野生株7404、HD-1-X和9510 不易获得并表达外源cry 基因,而高效野生株Bti. t-1897 的电转化频率较高,并获得以Btit-1897 为受体,对甜菜夜蛾、棉铃虫、黏虫和淡色库蚊均有毒力,具有cryIAc、cryIC、cryIV及cyt多价基因的广谱工程菌株.
【Abstract】 Several wild type Bacillus thuringiensis strains, which are highly toxic against Lepidopteran or Dipteran larvae, were detected for their cry gene types using PCR technology. Based on Southern hybridization, cry IAc gene was located on the 45MD plasmid of strain 7404, and cry IC gene was located on the 50MD plasmid of strain 9510. Conjugation and electroporation were used to genetically improve these strains. It showed that the wild type strains 7404, HD 1 X and 9510 were not easy to accept foreign cry gene by conjugation and eletroporation. While Bti. t 1897, which was highly toxic to Dipteran lavare, could be transformed at high freqency with the plasmid carring cry gene. Transformants Bti t 1897 T1~T14 and TT13 were obtained, which contained multi cry genes and exhibited toxicity to several Lepidopteran and Dipteran larvae.
【Key words】 Bacillus thuringiensis; cry gene lacolization; multi cry genes; conjugation; electroporation;
- 【文献出处】 南开大学学报(自然科学版) ,JOURNAL OF NANKAI UNIVERSITY , 编辑部邮箱 ,1999年03期
- 【分类号】S476.1
- 【被引频次】6
- 【下载频次】107