Cloning of the heat shock protein 60 gene from the stem borer, Chilo suppressalis, and analysis of expression characteristics under heat stress
1Institute of Applied Entomology, Yangzhou University, Yangzhou, 225009, China
2Department of Life Science, Fuyang Teachers College, Fuyang, 236032, China
3Department of Agriculture and Landscape Engineering, Xuzhou Higher Vocational School of Bioengineering, Xuzhou, 221006,
China
Abstract
Heat shock protein 60 is an important chaperonin. In this paper, hsp60 of the stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), was cloned by RT-PCR and rapid amplification of cDNA end (RACE) reactions. The full length cDNA of hsp60 consisted of 2142 bp, with an ORF of 1719 bp, encoding 572 amino acid residues, with a 5’UTR of 158 bp and a 3’UTR of 265 bp. Cluster analysis confirmed that the deduced amino acid sequence shared high identity with the reported sequences from other insects (77%-86%). To investigate whether hsp60 in C. suppressalis responds to thermal stress, the expression levels of hsp60 mRNA in larval haemocytes across temperature gradients from 31 to 390 C were analysed by real-time quantitative PCR. There was no significant difference for hsp60 expression from 28 to 310 C. The temperatures for maximal induction of hsp60 expression in haemocytes was close to 360 C. Hsp60 expression was observed by using flow cytometry. These results revealed that thermal stress significantly induced hsp60 expression and Hsp60 synthesis in larval haemocytes, and the expression profiles of Hsp60 at the mRNA and protein levels were in high agreement with each other from 33 to 390 C.
Keywords: Lepidoptera; Hsp60; RACE; haemocytes; temperature
Abbreviatons: Hsp, heat shock protein
Correspondence: a yzdu@yzu.edu.cn
Editor: David Schooley was editor of this paper
Received: 19 January 2010 | Accepted: 13 March 2010 | Published: 9 July 2010
Copyright: This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.
ISSN: 1536-2442 | Volume 10, Number 100
Cui YD, Du YZ, Lu MX, Qiang CK. 2010. Cloning of the heat shock protein 60 gene from the stem borer, Chilo suppressalis, and analysis of expression characteristics under heat stress. Journal of Insect Science 10:100, available online: insectscience.org/10.100
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