春尺蠖热激蛋白 AcinHsp70 基因的克隆、分子特性与表达分析.

[Objective] The purpose of the study was to clone heat shock protein Hsp70 gene from Apocheima cinerarius, analyze its physicochemical properties and expression, and explore the role of this gene in coping with the adverse environment of A. cinerarius. [Method] In the study, Hsp70 was cloned and named as AcinHsp70 (GenBank entry number: OP750249) based on the transcriptome data (under different low temperature stress of A. cinerarius larvae). The physical and chemical properties of Hsp70 gene were analyzed by using bioinformation analysis software. qPCR was used to detect the expression profiles of AcinHsp70 under different temperature stresses (10,5, 0, 5 and 25 °C) and under different time treatments at 4 °C. [Result] The total length of CDS of AcinHsp70 sequence was 1899 bp, encoding 633 amino acids, the molecular weight of protein was 69.91 kDa, and the isoelectric point was 5.51. The N-terminus of the protein lacked a signal peptide and transmembrane region. Phosphorylation sites were identified at serine, threonine and tyrosine residues (24, 26, and 8 sites respectively). No glycosylation sites were observed. The average hydrophilicity of the protein was 0.507, with a predicted instability coefficient of 33. 19, indicating its stability as a hydrophilic protein. Subcellular localization analysis revealed predominant cytoplasmic distribution. The results of sequence comparison and phylogenetic analysis showed that the A. cinerarius was closely related to Melitaea cinxia, and the sequence identity reached 92.91%. Gene expression results showed that AcinHsp70 expression was not significantly different under different temperature (except for 10 °C) stress, and the expression was significantly up-regulated after recovery. The expression level of the AcinHsp70 exhibited an initial increase followed by a subsequent decrease, with maximum expression after 1 hour treatment at 4 °C. [Conclusion] AcinHsp70 is closely related to the response of A. cinerarius to low temperature stress, and the results help to reveal the molecular function of Acinhsp70 under low temperature stress. [ABSTRACT FROM AUTHOR]