Document Detail

Stability of Cry1Ab protein during long-term storage for standardization of insect bioassays.
MedLine Citation:
PMID:  19833078     Owner:  NLM     Status:  MEDLINE    
The reliable use of purified Cry1Ab protein standards is a prerequisite for ecological studies and resistance monitoring programs of Cry1Ab-expressing transgenic corn. In this study the stability and activity of different Cry1Ab protein batches expressed in and purified from Escherichia coli were determined during two-year storage at different temperature conditions (4 degrees C, -20 degrees C, and -80 degrees C). SDS-Polyacrylamide gel electrophoresis showed degradation of the protein stored at 4 degrees C over four months, whereas no difference in the band intensity of the Cry1Ab proteins stored at -20 degrees C and -80 degrees C was observed. Bioassays with neonate larvae of Ostrinia nubilalis indicated that the biological activity of Cry1Ab varied from batch to batch, depending on the production process. Cry1Ab protein stored at 4 degrees C for four months showed a significantly decreasing activity measured as median lethal concentration (LC(50)), whereas the protein activity declined less than 11-fold after two years storage at -20 degrees C. When stored at -80 degrees C the toxin activity remained relatively stable for at least 30 months, as indicated by low LC(50) values of 7-10 ng Cry1Ab per cm(2) diet. These experiments demonstrate that appropriate long-term storage conditions of Cry1Ab protein standards are crucial for resistance monitoring programs of Bt corn, and storage at -80 degrees C is recommended.
Hang Thu Nguyen; Johannes A Jehle
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-10-16
Journal Detail:
Title:  Environmental biosafety research     Volume:  8     ISSN:  1635-7922     ISO Abbreviation:  Environ Biosafety Res     Publication Date:    2009 Apr-Jun
Date Detail:
Created Date:  2009-10-16     Completed Date:  2010-01-05     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101172622     Medline TA:  Environ Biosafety Res     Country:  France    
Other Details:
Languages:  eng     Pagination:  113-9     Citation Subset:  IM    
Agricultural Service Center Palatinate (DLR Rheinpfalz), Department of Phytopathology, Laboratory for Biotechnological Crop Protection, Neustadt an der Weinstrasse, Germany.
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MeSH Terms
Bacterial Proteins / chemistry*,  pharmacology
Biological Assay / methods*
Chemistry Techniques, Analytical / methods
Drug Stability
Endotoxins / chemistry*,  pharmacology
Hemolysin Proteins / chemistry*,  pharmacology
Insecticides / chemistry*,  pharmacology
Insects / drug effects*
Specimen Handling / methods
Reg. No./Substance:
0/Bacterial Proteins; 0/Endotoxins; 0/Hemolysin Proteins; 0/Insecticides; 0/insecticidal crystal protein, Bacillus Thuringiensis

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