Assessment of protoxin composition of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> strains by use of polyacrylamide gel block and mass spectrometry |
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Authors: | Zujiao Fu Yunjun Sun Liqiu Xia Xuezhi Ding Xiangtao Mo Xiaohui Li Kexue Huang Youming Zhang |
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Institution: | (1) College of Life Science, Hunan Normal University, Changsha, 410081, People’s Republic of China |
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Abstract: | Assessment of protoxin composition in Bacillus thuringiensis parasporal crystals is principally hampered by the fact that protoxins in a single strain usually possess high sequence homology.
Therefore, new strategies towards the identification of protoxins have been developed. Here, we established a powerful method
through embedding solubilized protoxins in a polyacrylamide gel block coupled to liquid chromatography-tandem mass spectrometry
(LC-MS/MS) analysis of in-gel-generated peptides for protoxin identification. Our model study revealed that four protoxins
(Cry1Aa, Cry1Ab, Cry1Ac and Cry2Aa) and six protoxins (Cry4Aa, Cry4Ba, Cry10Aa, Cry11Aa, Cyt1Aa, and Cyt2Ba) could be rapidly
identified from B. thuringiensis subsp. kurstaki HD1 and subsp. israelensis 4Q2-72, respectively. The experimental results indicated that our method is a straightforward tool for analyzing protoxin
expression profile in B. thuringiensis strains. Given its technical simplicity and sensitivity, our method might facilitate the present screening program for B. thuringiensis strains with new insecticidal properties.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Zujiao Fu and Yunjun Sun contributed equally to this work. |
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Keywords: | Bacillus thuringiensis Polyacrylamide gel block LC-MS/MS Protoxin expression profile Insecticidal activity |
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