Separation，Purification and Insecticidal Activity of cry8Ea2 Protein from Bacillus thuringiensis

doi:10.3969/j.issn.1000-7091.2012.06.011

Abstract

Abstract: cry8Ea2 protein is a kind of insecticidal crystal protein of Bacillus thuringiensis． In this study， cry8Ea2 was cloned into E． coli-B． thuringiensist shuttle vector pSXY422b， resulting from recombinant pScry8Ea2， and pScry8Ea2 was transformed into the Bt，acrystalliferous mutant generating HD8Ea2 engineering strain． cry8Ea2 protein was successfully expressed in HD8Ea2 stain． By anion exchange chromatography on molecular sieve chromatography， 1 30 kDa of toxin protein cry8Ea2 was purified． The activities of the cry8Ea2 toxin were evaluated by improved bioassays，and the LC50value to neonates of Holotrichia oblita and Holotrichia paralella was 0. 267 9 μg /mL and 0． 071 9 μg /mL， respectively． Insecticidal activity of cry8Ea2 to Holotrichia oblita was firstly reported in this study，which will broaden the protein insecticidal spectrum and provide new resources for genetically modified project．

Key words: Bacillus thuringiensisis, cry8, Shuttle vector, Protein purification, Bioassays

CLC Number:

Q786

WANG Wei, GUO Wei, XU Da-qing, SUN Wei-ming, LIU Da-qun, LI Rui-jun. Separation，Purification and Insecticidal Activity of cry8Ea2 Protein from Bacillus thuringiensis[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(6): 48-52. doi: 10.3969/j.issn.1000-7091.2012.06.011.

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http://www.hbnxb.net/EN/Y2012/V27/I6/48

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