Authors : Qiong Huang, Jie Hu, Lin Sun and Qin Wang
Abstract: Heat Shock/Stress Proteins (HSPs) are a group of stress proteins which are closely associated with organisms’ adaptability to environment and the heat shock protein 70 is the most conserved and important member. Researchers cloned an hsp70 gene from Tenebrio molitor larvae by PCR and RACE Method and determined the mRNA abundance in the beetle developmental stages by real-time qPCR. The cDNA cloned was 2,282 bp in full length containing a 115 bp 5'untranslated region rich in adenine, a 1,935 bp open reading frame and a 232 bp 3'untranslated regionrich in adenine and thymine. It also had seven repeats of the Heat Shock Element (HSE) nGAAn in its 5'UTR and a 22 bp Poly (A) tail in the 3'UTR. The deduced heat shock protein had a highly conserved N-terminal ATPase domain and a conserved C-terminal peptide-binding domain. The tertiary structure of ATPase domain was made of two large globular subdomains which were separated by a deep cleft and the peptide-binding domain was a sandwich of 2 four-stranded β-sheets with four loops protruding upwards and two α-helices. Real-time qPCR analysis revealed that the hsp70 mRNA expression in T. molitor was characterized by heat-inducible and developmental-regulation features. The results form a basis for further research on structure, function and expression regulation of HSPs from T. molitor as well as to decipher the relationship between HSPs and stress-resistance in the beetle.
Qiong Huang, Jie Hu, Lin Sun and Qin Wang, 2012. Cloning, Sequencing and Expression of a Heat Shock Protein 70 Gene from Tenebrio molitor. Journal of Animal and Veterinary Advances, 11: 4632-4643.