Constructing Bacillus thuringiensis strain that co-expresses Cry2Aa and chitinase

被引：10
|
作者：
Hu, Shengbiao [1 ]
Zhang, Xu [1 ]
Li, Yusheng [1 ]
Ding, Xuezhi [1 ]
Hu, Xiaofeng [1 ]
Yang, Qi [1 ]
Xia, Liqiu [1 ]
机构：
[1] Hunan Normal Univ, Coll Life Sci, Hunan Prov Key Lab Microbial Mol Biol, State Key Lab Breeding Base Microbial Mol Biol, Changsha, Hunan, Peoples R China
来源：
BIOTECHNOLOGY LETTERS | 2013年 / 35卷 / 07期
基金：
中国国家自然科学基金;
关键词：
Bacillus thuringiensis; Chitinase; Cry2Aa; Single-strand DNA; Triple recombineering; INSECTICIDAL ACTIVITY; GENE; RECOMBINATION; CLONING;
D O I：
10.1007/s10529-013-1171-0
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
A triple recombineering technique was used with plasmid pHT315 to produce pHTEC, a construct carrying chitinase and cry2Aa genes from Bacillus thuringiensis subsp. kurstaki 4.0718. Transformation of wild-type B. thuringiensis strain HD73 and the acrystalliferous strain Cry-B with pHTEC resulted in the recovery of recombinant strains that expressed Cry2Aa as cubic crystals in the cell pellet and soluble chitinase protein. The toxicity of HD73 (pHTEC) against Helicoverpa armigera larvae increased sevenfold when compared with HD73 (pHT315) harboring pHT315 vector. The triple recombineering protocol was optimized by comparing recombination efficacy mediated by RecE/RecT and Red alpha/Red beta and by using single-strand DNA as substrate.
引用
收藏
页码：1045 / 1051
页数：7
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