Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system
Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system
Eun-Kyung Jang(Korean Intellectual Property Office); 정병권(경북대학교); Gun-Seok Park(Kyungpook National University); Abdur Rahim Khan(Kyungpook National University); Sung-Jun Hong(Kyungpook National University); 박영준(경북대학교); WON-CHAN KIM(Kyungpook National University); JAE-HO SHIN(Kyungpook National University); Khalid M S Al-Ghamdi(King Abdulaziz University); Bassam Oudh Al-Johny(King Abdulaziz University); Yasir Anwar(King Abdulaziz University); Muhammad Faisal Siddiqui(Hazara University); Ihsan Ullah(King Abdulaziz University)
23권 1호, 172~176쪽
초록
Photorhabdus spp. has a high molecular weight Tc toxin with insecticidal activity. These toxins have been suggested as an alternative to BT toxin from Bacillus thuringiensis. Herein, we constructed a cosmid library with the genome of M1021 and screened the Escherichia coli clones showing insect toxicity by injecting each clone into Galleria mellonella larvae. In a total of 1020 clones, one clone with high insecticidal activity was selected and the nucleotide sequence of the cosmid of the clone was determined. In cosmid PtC28, a gene with 87% homology to the tccB gene of Photorhabdus temperata was found. Consequently, we have isolated the tccB gene cassette from the M1021 and expressed in E. coli expression vectors. The toxin was produced in the form of inclusion bodies but the denatured and refolded recombinant TccB showed strong mortality to the G. mellonella larvae.
Abstract
Photorhabdus spp. has a high molecular weight Tc toxin with insecticidal activity. These toxins have been suggested as an alternative to BT toxin from Bacillus thuringiensis. Herein, we constructed a cosmid library with the genome of M1021 and screened the Escherichia coli clones showing insect toxicity by injecting each clone into Galleria mellonella larvae. In a total of 1020 clones, one clone with high insecticidal activity was selected and the nucleotide sequence of the cosmid of the clone was determined. In cosmid PtC28, a gene with 87% homology to the tccB gene of Photorhabdus temperata was found. Consequently, we have isolated the tccB gene cassette from the M1021 and expressed in E. coli expression vectors. The toxin was produced in the form of inclusion bodies but the denatured and refolded recombinant TccB showed strong mortality to the G. mellonella larvae.
- 발행기관:
- 한국응용곤충학회
- 분류:
- 기타농생물