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Engineering the bacterial endophyte Burkholderia pyrrocinia JK-SH007 for the control of lepidoptera larvae by introducing the cry218 genes of Bacillus thuringiensis

Authors :
Choufei Wu
Liqin Zhang
Zhong Xing
Bingli Gao
Li Yang
Source :
Biotechnology & Biotechnological Equipment, Vol 31, Iss 6, Pp 1167-1172 (2017)
Publication Year :
2017
Publisher :
Informa UK Limited, 2017.

Abstract

To expand the usage of endophytes in agriculture and in forestry, the insecticidal gene cry218 of Bacillus thuringiensis was introduced into a poplar bacterial endophyte Burkholderia pyrrocinia JK-SH007. The cry218 gene was cloned by polymerase chain reaction (PCR) and was inserted into a PHKT2 expression vector that was introduced into the bacterial endophyte JK-SH007. By using sodium dodecyl sulphate polyacryl amide gel electrophoresis (SDS-PAGE) and western blotting, we confirmed that the engineered bacterial endophyte was successfully constructed, and it harboured insecticidal function after the bioassay in planta. The toxicity of the expressed insecticidal protein was analysed on second instar silkworm. The regression equation showed that the median lethal concentration (LC50) of the insecticidal protein was 0.77 (0.57–1.04) g/L at 72 h. The insecticidal bacteria genetically modified in this study have laid the foundation for further exploitation of biocontrol bacteria.

Details

ISSN :
13143530 and 13102818
Volume :
31
Database :
OpenAIRE
Journal :
Biotechnology & Biotechnological Equipment
Accession number :
edsair.doi.dedup.....f1b0a88e67d40028872507a6c3bf73f6