Cloning, characterization and expression of a new cry1Ab gene from Bacillus thuringiensisWB9

• Published in: Biotechnology letters Vol. 26; no. 20; p. 1557
• Main Authors: Huang, Zhipeng, Guan, Chunhong, Guan, Xiong
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Nature B.V 01-10-2004
• Subjects: Amino acids; Bacteria; Bacteriology; Cloning; E coli; Larvae; Proteins
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1023/B:BILE.0000045652.00137.1f

A new cry1Ab-type gene, cry1Ab17, 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^sub 33^ to Arg^sub 253^), II (Arg^sub 265^ to Phe^sub 462^), III (Asn^sub 464^ to Thr^sub 610^) of the Cry1Ab17 were 96.8%, 68.2% and 100% identical to the corresponding domains of Cry1Aa. Additionally, the cry1Ab17 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.[PUBLICATION ABSTRACT]