Cloning and Molecular Characterization of Hsp Genes from Anoplophora glabripennis and Their Responses to Cold Stress
Abstract
:1. Introduction
2. Results
2.1. Cloning and Sequence Analysis of AglaHsps
2.2. Multiple Sequence Alignment and Phylogenetic Analysis of AglaHsps
2.3. Prokaryotic Expression and Purification of AglaHsps
2.4. Antibody Titer Determination and Western Blotting Analysis
3. Discussion
3.1. The Diversified Structures of AglaHsps Underlie Their Functional Specificities
3.2. Evolutionary Trends and Environmental Adaptabilities of AglaHsps
3.3. AglaHsps Function in the Regulatory Networks
4. Materials and Methods
4.1. Insect Collection and Temperature Treatments
4.2. RNA Extraction and cDNA Library Construction
4.3. Cloning of AglaHsp Genes
4.4. Bioinformatic Analysis
4.5. Prokaryotic Expression and Protein Purification
4.6. Preparation of AglaHsps Antibody
4.7. Western Blotting Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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AglaHsp20.43 | AglaHsp71.18 | AglaHsp82.09 | AglaHsp89.76 | |
---|---|---|---|---|
Coding sequence | 546 bp | 1950 bp | 2154 bp | 2355 bp |
Amino acid residues | 181 aa | 649 aa | 717 aa | 784 aa |
Molecular weight (kDa) | 20.43 | 71.18 | 82.09 | 89.76 |
Theoretical isoelectric point | 6.50 | 5.52 | 5.00 | 4.88 |
Instability index | 44.10 | 37.03 | 38.39 | 40.06 |
Hydrophilic protein | −0.631 | −0.427 | −0.665 | −0.600 |
Transmembrane structures | No | No | No | No |
Signal peptide | No | No | No | aa 1-20 |
Subcellular localization | Cytoplasmic | Cytoplasmic | Cytoplasmic | Endoplasmic reticulum |
Alpha Helix | Extended Strand | Beta Turn | Random Coil | |
---|---|---|---|---|
AglaHsp20.43 | 24.86% | 16.02% | 5.52% | 53.59% |
AglaHsp71.18 | 41.29% | 18.80% | 7.40% | 32.51% |
AglaHsp82.09 | 48.81% | 15.76% | 5.86% | 29.57% |
AglaHsp89.76 | 51.02% | 13.78% | 3.70% | 31.51% |
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Xu, Y.; Li, Y.; Shi, F.; Zhang, S.; Zong, S.; Tao, J. Cloning and Molecular Characterization of Hsp Genes from Anoplophora glabripennis and Their Responses to Cold Stress. Int. J. Mol. Sci. 2022, 23, 11958. https://doi.org/10.3390/ijms231911958
Xu Y, Li Y, Shi F, Zhang S, Zong S, Tao J. Cloning and Molecular Characterization of Hsp Genes from Anoplophora glabripennis and Their Responses to Cold Stress. International Journal of Molecular Sciences. 2022; 23(19):11958. https://doi.org/10.3390/ijms231911958
Chicago/Turabian StyleXu, Yabei, Yurong Li, Fengming Shi, Sainan Zhang, Shixiang Zong, and Jing Tao. 2022. "Cloning and Molecular Characterization of Hsp Genes from Anoplophora glabripennis and Their Responses to Cold Stress" International Journal of Molecular Sciences 23, no. 19: 11958. https://doi.org/10.3390/ijms231911958