Latest Cover

Online Office

Contact Us

Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
Sponsored by:Sichuan Society of Zoologists; Chengdu Giant Panda Breeding Research Foundation; Sichuan Association of Wildlife Conservation; Sichuan University
Address:College of Life Sciences, Sichuan University, No.29, Wangjiang Road, Chengdu, Sichuan Province, 610064, China
Fax:+86-28-85410485 &
Your Position :Home->Past Journals Catalog->2015 Vol.34 No.4

Molecular Cloning of Antifreeze Protein Gene from Oodescelis chinensis (Coleoptera: Tenebrionidae) and Effect of Temperature on Its Expression
Author of the article:ZHANG Fengjuan, WANG Yan, WANG Lu, LI Zhaoyang, TAO Bolin, MA Ji
Author's Workplace:Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Sciences and Technology, Xinjiang University, Urumqi 830046, China
Key Words:Oodescelis chinensis; freeze tolerant insect; antifreeze protein; temperature stress; real-time fluorescence quantitative PCR
Abstract:Oodescelis chinensis (Coleoptera: Tenebrionidae), a rare freeze tolerant insect endemic to China, distributes in the desert and semi-desert regions of Xinjiang, China. However, it was unknown whether O. chinensis contained antifreeze protein gene. In this study, the total RNA of O.chinensis was extracted by using TRIzol reagent. Several sequences of antifreeze protein genes (Ocafp) were amplified by RT-PCR using the universal primers designed according to the homologous sequences of other beetles. The sequencing results showed that the deduced amino acid sequence of OcAFP shared the similar structure feature of other beetles containing the TCTXSXXCXXAX (where X stood for any amino acid) motif. However, there were many different mutations in the conserved TCT motifs, which may be due to the adaptation of O.chinensis to the extreme variation of environmental temperature since O.chinensis living in the soil surface. The results of real-time fluorescence quantitative PCR showed that the expression of Ocafp could be induced at-4 ℃, 5 ℃ and 42 ℃, respectively, but inhibited at 40 ℃. The results indicated that O. chinensis also contained antifreeze proteins, and the variability of the amino acid sequence was higher than that of other freeze avoidant insects. The expression of Ocafp could be induced either at low temperature or at high temperature, and this suggested that OcAFP may also have other functions.
2015,34(4): 518-523 收稿日期:2014-10-30
侯小娟, 梁娜, 邱立明, 等. 2013. 荒漠甲虫光滑鳖甲和小胸鳖甲抗冻蛋白基因及其3'-UTR的序列分析[J]. 应用昆虫学报, 50(5): 1219-1229.
李毅平, 龚和. 1998. 昆虫低温生物学: 昆虫耐冻的生理生化适应机制[J]. 昆虫知识, 35(6): 364-369.
刘忠渊, 王芸, 吕国栋, 等. 2006. Tenebrio molitor抗冻蛋白家族cDNA片段的克隆、序列分析及原核表达[J]. 遗传, 28(12): 1532-1540.
赵干, 马纪, 薛娜, 等. 2005. 新疆准噶尔小胸鳖甲抗冻蛋白基因的克隆和抗冻活性分析[J]. 昆虫学报, 48(5): 667-673.
Davies PL, Baardsnes J, Kuiper MJ, et al. 2002. Structure and function of antifreeze proteins[J]. Philosophical Transactions of the Royal Society of London Series B-Biological Sciences, 357: 927-935.
DeVries AL, Wohlschlag DE. 1969. Freezing resistance in some Antarctic fishes[J]. Science, 163(3871): 1073-1075.
Doucet D, Tyshenko MG, Davies PL, et al. 2002 A family of expressed antifreeze protein genes from the moth, Choristoneura fumiferana[J]. European Journal of Biochemistry, 269: 38-46.
Duman JG, Li N, Verleye D, et al. 1998. Molecular characterization and sequencing of antifreeze proteins from larvae of the beetle Dendroides canadensis[J]. Journal of Comparative Physiology B, 168: 225-232.
Graether SP, Kuiper MJ, Gagné SM, et al. 2000. Beta-helix structure and ice-binding properties of a hyperactive antifreeze protein from an insect[J]. Nature, 406(6793): 325-328.
Graham LA, Liou YC, Walker VK, et al. 1997. Hyperactive antifreeze protein from beetles[J]. Nature, 388(6644): 727-728.
Graham LA, Walker VK, Davies PL. 2000. Developmental and environmental regulation of antifreeze proteins in the mealworm beetle Tenebrio molitor[J]. European Journal of Biochemistry, 267(21): 6452-6458.
Hou F, Ma J, Liu X, et al. 2010. Seasonal changes in antifreeze protein gene transcription and water content of beetle Microdera punctipennis (Coleoptera, Tenebrionidae) from Gurbantonggut desert in central Asia[J]. Cryo Letters, 31(5): 359-370.
Lee RE, Denlinger DL. 1991. Insects at low temperature[M]. New York: Chapman and Hall: 513.
Leinala EK, Davies PL, Doucet D, et al. 2002. A beta-helical antifreeze protein isoform with increased activity[J]. Biological Chemistry, 277(36): 33349-33352.
Liou YC, Thibault P, Walker VK, et al. 1999. A complex family of highly heterogeneous and internally repetitive hyperactive antifreeze proteins from the beetle Tenebrio molitor[J]. Biochemistry, 38(35): 11415-11424.
Liu K, Jia ZC, Chen G, et al. 2005. Systematic size study of an insect antifreeze protein and its interaction with ice[J]. Biophysical Journal, 88(2): 953-958.
Marshall CB, Daley ME, Graham LA. 2002. Identification of the ice-binding face of antifreeze protein from Tenebrio molitor[J]. FEBS Letters, 529(2-3): 261-267.
Michaud MR, Denlinger DL. 2007. Shifts in carbohydrate, polyol, and amino acid pools during rapid cold hardening and diapause-associated cold hardening on flesh flies (Sarcophaga crassipalpis): a metabolomic comparison[J]. Journal of Comparative Physiology B, 177: 753-763.
Pedersen SA, Kristiansen E, Hansen BH, et al. 2006. Cold hardiness in relation to trace metal stress in the freeze-avoiding beetle Tenebrio molitor[J]. Journal of Insect Physiology, 52(8): 846-853.
Qiu L, Mao X, Hou F, et al. 2013. A novel function-thermal protective properties of an antifreeze protein from the summer desert beetle Microdera punctipennis[J]. Cryobiology, 66(1): 60-68.
Raymond JA, DeVries AL. 1977. Adsorption inhibition as a mechanism of freezing resistance in polar fishes[J]. Proceedings of the National Academy of Sciences of the United States of America, 74(6): 2589-2593.
Sejerkilde M, Sørensen JG, Loeschcke V. 2003. Effects of cold and heat hardening on thermal resistance in Drosophila melanogaster[J]. Journal of Insect Physiology, 49(8): 719-726.
CopyRight©2022 Editorial Office of Sichuan Journal of Zoology 蜀ICP备08107403号-3