Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system

Eun Kyung Jang; Byung Kwon Jung; Gun Seok Park; Abdur Rahim Khan; Sung Jun Hong; Yeong Jun Park; Won Chan Kim; Jae Ho Shin; Khalid M S Al-Ghamdi; Bassam Oudh Al-Johny; Yasir Anwar; Muhammad Faisal Siddiqui; Ihsan Ullah

doi:10.1016/j.aspen.2019.12.013

Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system

Eun Kyung Jang, Byung Kwon Jung, Gun Seok Park, Abdur Rahim Khan, Sung Jun Hong, Yeong Jun Park, Won Chan Kim, Jae Ho Shin, Khalid M S Al-Ghamdi, Bassam Oudh Al-Johny, Yasir Anwar, Muhammad Faisal Siddiqui, Ihsan Ullah

School of Applied Bioscience

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

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.

Original language	English
Pages (from-to)	172-176
Number of pages	5
Journal	Journal of Asia-Pacific Entomology
Volume	23
Issue number	1
DOIs	https://doi.org/10.1016/j.aspen.2019.12.013
State	Published - Apr 2020

Keywords

Galleria mellonella
M1021
pET21a(+)
tccB gene
Toxin complexes

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Jang, E. K., Kwon Jung, B., Park, G. S., Rahim Khan, A., Hong, S. J., Park, Y. J., Kim, W. C., Shin, J. H., M S Al-Ghamdi, K., Oudh Al-Johny, B., Anwar, Y., Faisal Siddiqui, M., & Ullah, I. (2020). Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system. Journal of Asia-Pacific Entomology, 23(1), 172-176. https://doi.org/10.1016/j.aspen.2019.12.013

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title = "Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system",

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.",

keywords = "Galleria mellonella, M1021, pET21a(+), tccB gene, Toxin complexes",

author = "Jang, {Eun Kyung} and {Kwon Jung}, Byung and Park, {Gun Seok} and {Rahim Khan}, Abdur and Hong, {Sung Jun} and Park, {Yeong Jun} and Kim, {Won Chan} and Shin, {Jae Ho} and {M S Al-Ghamdi}, Khalid and {Oudh Al-Johny}, Bassam and Yasir Anwar and {Faisal Siddiqui}, Muhammad and Ihsan Ullah",

note = "Publisher Copyright: {\textcopyright} 2019 Korean Society of Applied Entomology",

year = "2020",

month = apr,

doi = "10.1016/j.aspen.2019.12.013",

language = "English",

volume = "23",

pages = "172--176",

journal = "Journal of Asia-Pacific Entomology",

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Jang, EK, Kwon Jung, B, Park, GS, Rahim Khan, A, Hong, SJ, Park, YJ, Kim, WC , Shin, JH, M S Al-Ghamdi, K, Oudh Al-Johny, B, Anwar, Y, Faisal Siddiqui, M & Ullah, I 2020, 'Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system', Journal of Asia-Pacific Entomology, vol. 23, no. 1, pp. 172-176. https://doi.org/10.1016/j.aspen.2019.12.013

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T1 - Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system

AU - Jang, Eun Kyung

AU - Kwon Jung, Byung

AU - Park, Gun Seok

AU - Rahim Khan, Abdur

AU - Hong, Sung Jun

AU - Park, Yeong Jun

AU - Kim, Won Chan

AU - Shin, Jae Ho

AU - M S Al-Ghamdi, Khalid

AU - Oudh Al-Johny, Bassam

AU - Anwar, Yasir

AU - Faisal Siddiqui, Muhammad

AU - Ullah, Ihsan

PY - 2020/4

Y1 - 2020/4

N2 - 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.

AB - 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.

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ER -

Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system

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Keywords

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