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Toxins 2014, 6(7), 2024-2040; doi:10.3390/toxins6072024
Article

Identification and Characterization of the Insecticidal Toxin “Makes Caterpillars Floppy” in Photorhabdus temperata M1021 Using a Cosmid Library

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Received: 19 April 2014; in revised form: 20 June 2014 / Accepted: 2 July 2014 / Published: 10 July 2014
(This article belongs to the Section Bacterial Toxins)
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Abstract: Photorhabdus temperata is an entomopathogenic enterobacterium; it is a nematode symbiont that possesses pathogenicity islands involved in insect virulence. Herein, we constructed a P. temperata M1021 cosmid library in Escherichia coli XL1-Blue MRF` and obtained 7.14 × 105 clones. However, only 1020 physiologically active clones were screened for insect virulence factors by injection of each E. coli cosmid clone into Galleria mellonella and Tenebrio molitor larvae. A single cosmid clone, PtC1015, was consequently selected due to its characteristic virulent properties, e.g., loss of body turgor followed by death of larvae when the clone was injected into the hemocoel. The sequence alignment against the available sequences in Swiss-Prot and NCBI databases, confirmed the presence of the mcf gene homolog in the genome of P. temperata M1021 showing 85% homology and 98% query coverage with the P. luminescens counterpart. Furthermore, a 2932 amino acid long Mcf protein revealed limited similarity with three protein domains. The N-terminus of the Mcf encompassed consensus sequence for a BH3 domain, the central region revealed similarity to toxin B, and the C-terminus of Mcf revealed similarity to the bacterial export domain of ApxIVA, an RTX-like toxin. In short, the Mcf toxin is likely to play a role in the elimination of insect pests, making it a promising model for use in the agricultural field.
Keywords: insecticidal toxin; Galleria mellonella; makes caterpillar floppy; Photorhabdus temperata; Tenebrio molitor insecticidal toxin; Galleria mellonella; makes caterpillar floppy; Photorhabdus temperata; Tenebrio molitor
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Ullah, I.; Jang, E.-K.; Kim, M.-S.; Shin, J.-H.; Park, G.-S.; Khan, A.R.; Hong, S.-J.; Jung, B.-K.; Choi, J.; Park, Y.; Kwak, Y.; Shin, J.-H. Identification and Characterization of the Insecticidal Toxin “Makes Caterpillars Floppy” in Photorhabdus temperata M1021 Using a Cosmid Library. Toxins 2014, 6, 2024-2040.

AMA Style

Ullah I, Jang E-K, Kim M-S, Shin J-H, Park G-S, Khan AR, Hong S-J, Jung B-K, Choi J, Park Y, Kwak Y, Shin J-H. Identification and Characterization of the Insecticidal Toxin “Makes Caterpillars Floppy” in Photorhabdus temperata M1021 Using a Cosmid Library. Toxins. 2014; 6(7):2024-2040.

Chicago/Turabian Style

Ullah, Ihsan; Jang, Eun-Kyung; Kim, Min-Sung; Shin, Jin-Ho; Park, Gun-Seok; Khan, Abdur R.; Hong, Sung-Jun; Jung, Byung-Kwon; Choi, JungBae; Park, YeongJun; Kwak, Yunyoung; Shin, Jae-Ho. 2014. "Identification and Characterization of the Insecticidal Toxin “Makes Caterpillars Floppy” in Photorhabdus temperata M1021 Using a Cosmid Library." Toxins 6, no. 7: 2024-2040.


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