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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
- 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.
- Subjects :
- 0301 basic medicine
Larva
Expression vector
biology
lcsh:Biotechnology
fungi
Bacillus thuringiensis
biology.organism_classification
Endophyte
insecticidal activity
Microbiology
Lepidoptera genitalia
03 medical and health sciences
030104 developmental biology
Burkholderia pyrrocinia
lcsh:TP248.13-248.65
Botany
expression vector
Gene
cry218 gene
Biotechnology
Subjects
Details
- ISSN :
- 13143530 and 13102818
- Volume :
- 31
- Database :
- OpenAIRE
- Journal :
- Biotechnology & Biotechnological Equipment
- Accession number :
- edsair.doi.dedup.....f1b0a88e67d40028872507a6c3bf73f6