Molecular Nutrition

Cloning, Sequence Analysis and Expression Profile of Copper/Zinc Superoxide Dismutase Gene of Apis cerana cerana

  • LIU Junfeng ,
  • LIU Tingting ,
  • WANG Huan ,
  • WANG Zilong ,
  • YAN Weiyu ,
  • ZENG Zhijiang ,
  • WU Xiaobo
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  • Honeybee Research Institute, Jiangxi Agricultural University, Nanchang 330045, China

Received date: 2012-02-29

  Online published: 2012-08-03

Abstract

This experiment was conducted to clone the cDNA and study the expression profile of copper/zinc superoxide dismutase (SOD1) gene of Apis cerana cerana. The SOD1 gene of Apis cerana cerana was cloned by nested PCR. The expression level of SOD1 gene in different tissues (brain, thorax and abdomen of adults) and at different developmental stages (3- and 6-day-old larva instars, 1- and 4-day-old pupas and 1- and 7-day-old adults) of Apis cerana cerana was detected using quantitative real-time PCR (qRT-PCR). The results showed that the full length of cDNA of SOD1 gene of Apis cerana cerana (GenBank accession number is JN700517) was 631 bp, encoding 152 amino acids, and the predicted molecular weight and isoelectric point were 15.65 ku and 6.21, respectively. The deduced amino acid sequence of SOD1 of Apis cerana cerana showed the highest similarity (99%) with that of Apis mellifera ligustica, and had 67%~85% similarity with other insects (such as Drosophila melanogaster, Anopheles gambiae, Spodoptera exigua, Bombyx mori, Bombus ignitus and so on). The expression level of SOD1 gene was significantly different at different developmental stages of Apis cerana cerana, and the peak appeared in 6-day-old larva instar, while the lowest value appeared in 4-day-old pupa. The expression level in the brain and abdomen was significantly higher than that in the thorax (P<0.05). The cDNA of SOD1 gene of Apis cerana cerana is successfully cloned in this study, and the full length is 631 bp, encoding 152 amino acids. The expression profile at different developmental stages and in different tissues is different.

Cite this article

LIU Junfeng , LIU Tingting , WANG Huan , WANG Zilong , YAN Weiyu , ZENG Zhijiang , WU Xiaobo . Cloning, Sequence Analysis and Expression Profile of Copper/Zinc Superoxide Dismutase Gene of Apis cerana cerana[J]. Chinese Journal of Animal Nutrition, 2012 , 24(8) : 1512 -1519 . DOI: 10.3969/j.issn.1006-267x.2012.08.015

References

[1] PARKER J D,PARKER K M,KEER L.Molecular phylogenetic evidence for an extracellular Cu Zn superoxide dismutase gene in insects[J].Insect Molecular Biology,2004,13(6):587-594.  

[2] 曾志将.养蜂学[M].2版.北京:中国农业出版社,2009.

[3] 李红亮,聂文敏,高其康,等.中华蜜蜂气味结合蛋白ASP2 cDNA的克隆及原核表达[J].中国农业科学,2008,41(3):933-938.

[4] CHOI Y S,LEE K S,YOON H J,et al.Bombus ignitus Cu,Zn superoxide dismutase (SOD1):cDNA cloning,gene structure,and up-regulation in response to paraquat,temperature stress,or lipopolysaccharide stimulation[J].Comparative Biochemistry and Physiology,Part B:Biochemistry and Molecular Biology,2006,144 (3) 365-371.

[5] WEIRICH G F,COLLINS A M,WILLIAMS V P.Antioxidant enzymes in the honey bee (Apis mellifera)[J].Apidologie,2002,33:3-14.

[6] CORONA M,HUGHES K A,WEAVER D B,et al.Gene expression patterns associated with queen honey bee longevity[J].Mechanisms of Ageing and Development,2005,126:1230-1238

[7] 肖培新,吴在富,刘昭华,等.意大利工蜂不同发育时期抗氧化酶基因mRNA表达量的变化[J].昆虫学报,2010,53(11):1202-1206.

[8] 丛海峰,徐世清,司马杨虎,等.野桑蚕铜锌超氧化物歧化酶基因的克隆分析与原核表达[J].蚕业科学,2007,33(2):234-240.

[9] 姚立虎,朱保建,刘朝良,等.柞蚕铜锌超氧化物歧化酶基因的克隆与表达分析[J].蚕业科学,2009,35(3):547-551.

[10] 蒋芳平,应琼琼,顾蔚.对羟基苯甲酸丙酯对果蝇超氧化物歧化酶(SOD)基因表达的影响[J].食品科学,2011,32(19):170-173.

[11] LIU W H,SAINT D A.A new quantitative method of real time reverse transcription polymerase chain reaction assay based on simulation of polymerase chain reaction kinetics[J].Analytical Biochemistry,2002,302(1):52-59.  

[12] THOMPSON J D,GIBSON T J,PLEWNIAK F,et al.The CLUSTAL_X windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools[J].Nucleic Acids Research,1997,25(24):4876-4882.  

[13] TAMURA K,DUDLEY J,NEI M,et al.MEGA4:molecular evolutionary genetics analysis (MEGA) software version 4.0[J].Molecular Biology and Evolution,2007,24:1596-1599.

[14] ARHONTAKI K,ELIOPOULOS E,GOULIELMOS G,et al.Functional constraints of the Cu,Zn superoxide dismutase in species of the Drosophila melanogaster subgroup and phylogenetic analysis[J].Journal of Molecular Evolution,2002,55(6):745-756.  

[15] 陈永,龚亮,左洪亮,等.斜纹夜蛾线粒体复合物ⅢFe-S蛋白基因克隆、序列分析及在不同发育阶段的表达特征[J].昆虫学报,2011,54(7):762-768.

[16] BOWLER C,MONTAGU M V,INZE D.Superoxide dismutase and stress tolerance[J].Annual Review Plant Physiology and Plant Molecular Biology,1992,43:83-116.

[17] 刘光楠,张飞,颜伟玉,等.蜂王幼虫与工蜂幼虫发育期食物消耗量的研究[J].应用昆虫学报,2011,48(1):113-115.

[18] 冯毛,李建科.王浆高产蜜蜂和原种意大利蜜蜂咽下腺发育蛋白质组分析[J].中国农业科学,2009,42(2):677-687.

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