Cloning of cry8Fa2 Gene and Its Expression in Bacillus thuringiensis Acrystalliferous Mutant Strain

doi:10.7668/hbnxb.2011.03.004

Abstract

Abstract: The 3.5 kb fragments was amplified by PCR using a pair of Bt cry8-type genes special primers,JJX5 and JJX3,and inserted into vector pMD 18-T,the new recombinant plasmid,pMD-cry8new,was isolated and obtained.Nucleic acid sequence analysis showed that this gene was 3 525 base pairs encoding 1 174 amino acids,which were homolog of 99.8% compared with Cry8Fal,the molecular weight of the protein was 133.05 kDa with isoelectric point pH 4.69.This gene sequence had been registered in GenBank(accession number was HQ174208),and named Cry8Fa2 as a novel gene by Bacillus thuringiensis Delta Endotoxin Nomenclature Committee.The cry8Fa2 gene could be formed parasporal crystal and expressed as a 130 kDa protein in Bt acrystalliferous mutant strain HD-73-.Bioassay result showed the expression product of cry8Fa2 gene was not toxic to the larvae of Holotrichia parallela and H.oblita.

Key words: Bacillus thuringiensis, cry8Fa2 gene, Gene cloning, Protein expression

CLC Number:

ZANG Da-kang, ZHENG Gui-ling, ZHOU Hong-xu, ZHANG Yuan, LI Guo-xun, LI Chang-you. Cloning of cry8Fa2 Gene and Its Expression in Bacillus thuringiensis Acrystalliferous Mutant Strain[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(3): 16-20. doi: 10.7668/hbnxb.2011.03.004.

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