title: Expression profiles of the heat shock protein 70 gene in response to heat stress in Agrotis c-nigrum (Lepidoptera: Noctuidae).
abstract: Heat shock proteins (HSPs) are molecular chaperones, and their overexpression enhances the survivability and stress tolerance of the cell. To understand the characteristics of HSP70 in Agrotis c-nigrum Linnaeus larvae, the coding sequence of this protein was cloned, and the effect of heat stress on transcription and protein properties was assessed. The obtained cDNA sequence of HSP70 was 2,213 bp, which contained an ORF of 1,965 bp and encoded 654 amino acid residues. Isolated HSP70 cDNA demonstrated more than 80% identity with the sequences of other known insect HSP70s. Next, HSP70 was expressed in Escherichia coli BL21 (DE3) cells and identified using SDS-PAGE and western blotting analyses. In addition, anti-HSP70-specific antisera were prepared using a recombinant HSP70 protein, and the results showed that this antisera was very specific to AcHSP70. Real-time quantitative polymerase chain reaction detected the relative transcription of the HSP70 gene in larvae and the transcription of A. c-nigrum in response to high temperatures. Induction of HSP70 was up-regulated to peak expression at 36°C.
authors: College of Agriculture, Northeast Agricultural University, Harbin 150030, China.
authors: Ling
authors: L
authors: Wang
authors: College of Agriculture, Northeast Agricultural University, Harbin 150030, China Virus-free Seedling Research Institute, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China.
authors: Shuai
authors: S
authors: Yang
authors: College of Agriculture, Northeast Agricultural University, Harbin 150030, China kjzhao@163.com.
authors: Kuijun
authors: K
authors: Zhao
authors: College of Agriculture, Northeast Agricultural University, Harbin 150030, China.
authors: Lanlan
authors: L
authors: Han
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000502:physiology; D020869:Gene Expression Profiling; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006358:Hot Temperature; D007814:Larva; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D018076:DNA, Complementary; D018840:HSP70 Heat-Shock Proteins
keywords: Agrotis c-nigrum Linnaeus; expression profile; heat shock protein 70; heat stress