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Cloning, Expression of cry1Aa21 Gene and Its Activity to Brown Planthopper Activity

Author: LinHongLi
Tutor: GuanXiong
School: Fujian Agriculture and Forestry University
Course: Pesticides
Keywords: Bacillus thuringiensis Cry1Aa21 protein Brown planthopper activity
CLC: S476.1
Type: Master's thesis
Year: 2011
Downloads: 24
Quote: 0
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Abstract


In order to of Bacillus thuringiensis ( Bacillus thuringiensis , referred to as the Bt ) standard strains of The HD - 1 total DNA as a template, through the amplification of cry1A gene to those PCR method it to connect , will the PCR product is recovered by with the the pMD18-T vector , to build clone recombinant bacteria pMD18T - cry1A from , by sequencing and analysis of the gene for the complete ORF 3531 bp , encoding a protein of 1176 amino acids , molecular weight 133128Da , isoelectric point of 5.02 . NCBI BLAST software analysis of its nucleotide sequence , the results show this gene cry1Aa1 cry1Ac1 have 99 % homology . The Nomenclature Committee named by the Bt new gene cry1Aa21 , the NCBI Login JF699284 . The of the cry1Aa21 gene pET32a expression vector to construct a recombinant strain BL21-pET32a-cry1Aa21 . SDS-PAGE analysis showed that the target protein of about 150 kDa , which contains about 20 kDa HIS pins . Extraction Cry1Aa21 protein to uninduced protein and the original strain HD - 1 crystal protein as control , research on the biological activity of the diamondback moth Plutella xylostella ( L. ) and brown planthopper Nilaparvata lugens ( Stal ) , results found Cry1Aa21 protein and HD - 1 crystal protein on the diamondback moth 48-hour LC50 of 2.429μg/mL and 0.691μg/mL . The use of artificial feed membrane fed method, the to carried out the Cry1A protein and the the activity of validation of of the Bt crystal protein in the within 72 hours of on the brown planthopper . In series of concentrations of the Cry1Aa21 of protein 12700μg/mL to 0.0127μg/mL , HD - 1 In series concentration 15900μg/mL 0.0159μg/mL , are the brown planthopper 4,5 instar nymphs and adult mortality rates had no significant effect . Cry1Aa21 and HD - 1 protein activated by trypsin the brown planthopper 4,5 age and adults had no significant effect .

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CLC: > Agricultural Sciences > Plant Protection > A variety of control methods > Biological control > The use of microbial pathogens
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