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抗光肩星天牛苏云金芽胞杆菌Bt886菌株的分离及对毒蛋白编码基因的初步鉴定
引用本文:陈军,杜孟芳,尹新明,王学聘,卢孟柱. 抗光肩星天牛苏云金芽胞杆菌Bt886菌株的分离及对毒蛋白编码基因的初步鉴定[J]. 林业科学, 2004, 40(5): 138-142
作者姓名:陈军  杜孟芳  尹新明  王学聘  卢孟柱
作者单位:中国林业科学研究院林业研究所,国家林业局林木培育重点实验室,北京,100091;河南农业大学植物保护学院,郑州,450002
基金项目:国家重点基础研究规划项目 (TG19990 160 0 0 )资助。
摘    要:从拟谷盗虫尸中分离到一株对鞘翅目害虫光肩星天牛幼虫毒杀作用明显的Bt菌株Bt886。幼虫死亡率高达 6 0 % ,存活者生长明显受到抑制。经过晶体形态特征和杀虫谱分析 ,初步确认该菌株属于cry3类。比较cry3类基因的保守序列 ,利用一对cry3特异引物成功地从Bt886菌株中克隆到了一段基因片段并克隆至pGEM -T载体上。测序分析发现该片段与cry3Aa1基因具有很高的同源性 ,确定来自毒蛋白编码基因。Southern杂交分析表明该基因位于该菌株的质粒上。

关 键 词:天牛  Bt菌  cry3A基因  Southern杂交分析
文章编号:1001-7488(2004)05-0138-05
修稿时间:2004-03-09

Selection of Bacillus thuringiensis Strain against the Longhorned Beetles and Preliminary Characterization of Its Insecticidal Gene
Chen Jun ) Wang Xuepin ) Lu Mengzhu ). Selection of Bacillus thuringiensis Strain against the Longhorned Beetles and Preliminary Characterization of Its Insecticidal Gene[J]. Scientia Silvae Sinicae, 2004, 40(5): 138-142
Authors:Chen Jun ) Wang Xuepin ) Lu Mengzhu )
Affiliation:Chen Jun 1) Wang Xuepin 4) Lu Mengzhu 5)
Abstract:A Bacillus thuringiensis strain Bt886 toxic to Longhorned Beetle (Anoplophora glabripennis) larvae were isolated from dead body of Tribolium macleby. The larvae fed with Bt886 exhibited high mortality (60%) and significant delay in growth. The shape of its crystal and spectrum of affected insects indicated that Bt886 could harbor cry3 like genes. A pair of primers was designed based on the conservative region of cry3 genes and a DNA fragment was successfully obtained by amplification. The fragment was further cloned into pGEM-T and the sequence analysis showed highly homologous to cry3 Aa1 gene with only one nucleotide difference. The fragment was located on plasmid of Bt886 by Southern analysis.
Keywords:Longhorned Beetle   Bacillus thuringiensis   cry 3A gene   Southern analysis
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