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苏云金芽孢杆菌以色列亚种δ-内毒素DNA在球形芽孢杆菌中的克隆与表达

Cloning and expression of Bacillus thuringiensis israelensis delta-endotoxin DNA in B. sphaericus.

作者信息

Bar E, Lieman-Hurwitz J, Rahamim E, Keynan A, Sandler N

机构信息

Department of Biological Chemistry, Hebrew University of Jerusalem, Israel.

出版信息

J Invertebr Pathol. 1991 Mar;57(2):149-58. doi: 10.1016/0022-2011(91)90110-c.

Abstract

Bacillus thuringiensis israelensis delta-endotoxin genes were cloned into Bacillus sphaericus 2362, producing stable transformants reacting with antibody to the 28- and 65-kDa B. thuringiensis israelensis crystal proteins and approximately 10 times more toxic to Aedes mosquito larvae than the original host strain. The LC50 after 48 hr of exposure of Aedes larvae to the most active transformed clone was 0.19 microgram/ml, compared with an LC50 of 1.9 microgram/ml for B. sphaericus 2362 and less than 0.1 microgram/ml for B. thuringiensis israelensis. The cloning vector, plasmid pPL603E, was also effective in transforming B. subtilis 1E20 with B. thuringiensis israelensis DNA, producing highly toxic clones with less stable gene expression than the clones of B. sphaericus.

摘要

苏云金芽孢杆菌以色列变种的δ-内毒素基因被克隆到球形芽孢杆菌2362中,产生了稳定的转化体,这些转化体与针对28 kDa和65 kDa苏云金芽孢杆菌以色列变种晶体蛋白的抗体发生反应,并且对伊蚊幼虫的毒性比原始宿主菌株高约10倍。将伊蚊幼虫暴露于最具活性的转化克隆48小时后的半数致死浓度(LC50)为0.19微克/毫升,相比之下,球形芽孢杆菌2362的LC50为1.9微克/毫升,而苏云金芽孢杆菌以色列变种的LC50小于0.1微克/毫升。克隆载体质粒pPL603E在用苏云金芽孢杆菌以色列变种DNA转化枯草芽孢杆菌1E20时也有效,产生了毒性很高但基因表达稳定性不如球形芽孢杆菌克隆的克隆体。

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