Cloning, characterization and expression of a new cry 1Ab gene from Bacillus thuringiensis WB9.
A new cry 1Ab-type gene, cry 1Ab17, was cloned from Bacillus thuringiensis WB9 by PCR. Nucleotide sequence indicated that the open reading frames (ORFs) consists of 3471 bases and encodes a protein of 1156 residues with a calculated molecular weight of 130.5 kDa and an pI value of 5.04. Homology comparison revealed that the deduced amino acid sequence of Cry1Ab17 had 95.4% to 99.7% identity with those of the known Cry1Ab proteins. The Cry1Ab17 was one residue longer than the known Cry1Ab (except for Cry1Ab2). Domain I (Tyr(33) to Arg(253)), II (Arg(265) to Phe(462)), III (Asn(464) to Thr(610)) of the Cry1Ab17 were 96.8%, 68.2% and 100% identical to the corresponding domains of Cry1Aa. Additionally, the cry 1Ab17 gene was expressed in Escherichia coli BL21 under the control of T7 promoter and the Cry1Ab17 isolated from the culture medium was toxic to 3rd instar Plutella xylostella larvae.
Huang
Zhipeng
Z
Key Laboratory of Biopesticide and Chemical Biology, and Biotechnology Center, College of Life Science, Fujian Agriculture & Forestry University, Ministry of Education, Fuzhou, Fujian 350002, P.R. China. zhipengh6841@yahoo.com.cn
Guan
Chunhong
C
Guan
Xiong
X
D000595:Amino Acid Sequence; D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics* / Q000378:metabolism*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000494:pharmacology; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000737:chemistry* / Q000494:pharmacology; D003001:Cloning, Molecular / Q000379:methods*; D004731:Endotoxins / Q000096:biosynthesis* / Q000737:chemistry* / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006460:Hemolysin Proteins; D007915:Lepidoptera / Q000187:drug effects; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D016019:Survival Analysis
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D011994:Recombinant Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis