王姿曼,郝瑞娟,邓岳文,李俊辉

• 1:广东海洋大学水产学院
• 2:广东省珍珠养殖与加工工程技术研究中心

摘要(Abstract)：

【目的】克隆马氏珠母贝(Pinctada martensii)法尼酸甲基转移酶(FAMe T)基因,并分析其在各组织中的表达。【方法】利用cDNA末端快速扩增技术(RACE)克隆获得马氏珠母贝FAMeT(PmFAMeT)基因的cDNA全长序列,利用实时荧光定量PCR(qPCR)方法分析PmFAMeT在马氏珠母贝不同组织中的表达模式。【结果与结论】PmFAMeT包含5′非编码区120 bp,3′非编码区968 bp和开放阅读框(ORF)1 536 bp,编码511个氨基酸。序列分析表明,PmFAMeT含有信号肽序列和跨膜结构域,并有WSC结构域、TSP1结构域和2个Methyltransf_FA结构域。将推导的PmFAMeT氨基酸序列与其他物种的FAMeT序列进行比对发现,不同物种的Methyltransf_FA序列同源性较高。PmFAMeT在马氏珠母贝的各个组织中均有表达,且在边缘膜、肝胰腺和性腺中的表达量显著高于其他组织。

关键词(KeyWords)： 马氏珠母贝;法尼酸甲基转移酶;基因克隆;表达模式

Abstract:

Keywords:

基金项目(Foundation): 农业产业体系专项(CARS-49);; 广东省自然科学基金项目(2017A030307024);; 广东省省级科技计划项目(2017A030303076);; 广东省渔港建设与渔业发展专项(B201601-Z09)

作者(Author): 王姿曼,郝瑞娟,邓岳文,李俊辉

参考文献(References)：

• [1]GUNAWARDENE Y,TOBE S,BENDENA W,et al.Function and cellular localization of farnesoic acid O-methyltransferase(FAMeT)in the shrimp,Metapenaeus ensis[J].European Journal of Biochemistry,2002,269(14):3587-3595.
• [2]NAGARAJU G.Is Methyl farnesoate a crustacean hormone?[J].Aquaculture,2007,272(1-4):39-54.
• [3]LAUFER H,DEMIR N,PAN X J,et al.Methyl farnesoate controls adult male morphogenesis in the crayfish,Procambarus clarkii[J].Journal of Insect Physiology,2005,51(4):379-384.
• [4]RODRIGUEZ E M,GRECO L,MEDESANI D A,et al.Effect of methyl farnesoate,alone and in combination with other hormones,on ovarian growth of the red swamp crayfish,Procambarus clarkii,during vitellogenesis[J].General and Comparative Endocrinology,2002,125(1):34-40.
• [5]TAMONE S L,CHANG E S.Methyl farnesoate stimulates ecdysteroid secretion from crab Y-organs in vitro[J].General and Comparative Endocrinology,1993,89(3):425-432.
• [6]HUI J,TOBE S,CHAN S M.Characterization of the putative farnesoic acid O-methyltransferase(LvFAMeT)c DNA from white shrimp,Litopenaeus vannamei:evidence for its role in molting[J].Peptides,2008,29(2):252-260.
• [7]谢熙,朱冬发,崔晓雨,等.三疣梭子蟹FAMeT基因克隆及其在蜕皮周期中的表达水平[J].水产学报,2013,37(7):994-1001.
• [8]谢熙.三疣梭子蟹卵巢发育过程中FAMeT和MIH基因表达变化[D].宁波:宁波大学,2013.
• [9]夏西超.Kk-42对凡纳滨对虾生长发育相关基因时空表达的影响[D].新乡:河南师范大学,2011.
• [10]DU X D,FAN G Y,JIAO Y,et al.The pearl oyster Pinctada fucata martensii genome and multi-omic analyses provide insights into biomineralization[J].Giga Science,2017,6(8):1-12.
• [11]WANG Q H,HAO R J,ZHAO X X,et al.Identification of EGFR in pearl oyster(Pinctada fucata martensii)and correlation analysis of its expression and growth traits[J].Bioscience Biotechnology and Biochemistry,2018,82(7):1073-1080.
• [12]HAO R J,ZHENG Z,WANG Q H,et al.Molecular and functional analysis of PmCHST1b in nacre formation of Pinctada fucata martensii[J].Comparative Biochemistry and Physiology.Part B,Biochemistry&Molecular Biology,2018,225:13-20.
• [13]ZHENG Z,LIANG J L,HUANG R L,et al.Identification of a novel miR-146a from Pinctada martensii involved in the regulation of the inflammatory response[J].Fish&Shellfish Immunology,2016,54:40-45.
• [14]HUANG J L,LI S G,LIU Y J,et al.Hemocytes in the extrapallial space of Pinctada fucata are involved in immunity and biomineralization[J].Scientific Reports,2018,8(1):4657.DOI:10.1038/s41598-018-22961-y
• [15]郝瑞娟,郑哲,王庆恒,等.马氏珠母贝磺基转移酶Pm CHST9基因的分子特征与表达分析[J].基因组学与应用生物学,2015,34(11):2387-2394.
• [16]TONG S M,CHEN Y,ZHU J,et al.Subcellular localization of five singular WSC domain-containing proteins and their roles in beauveria bassiana responses to stress cues and metal ions[J].Environmental Microbiology Reports,2016,8(2):295-304.
• [17]ADAMS J.Thrombospondins:multifunctional regulators of cell interactions[J].Annual Review of Cell and Developmental Biology,2001,17:25-51.
• [18]LAWLER J,HYNES R.The Structure of human thrombospondin,an adhesive glycoprotein with multiple calcium-binding sites and homologies with several different proteins[J].The Journal of Cell Biology,1986,103(5):1635-1648.
• [19]SMITH K F,NOLAN K F,REID K B,et al.Neutron and X-ray scattering studies on the human complement protein properdin provide an analysis of the thrombospondin repeat[J].Biochemistry,1991,30(32):8000-8008.
• [20]VIEIRA C U,BONETTI A M,SIMOES Z L P,et al.Farnesoic acid O-methyl transferase(FAMeT)isoforms:conserved traits and gene expression patterns related to caste differentiation in the stingless bee,Melipona Scutellaris[J].Archives of Insect Biochemistry and Physiology,2008,67(2):97-106.
• [21]NAGARAJU G,REDDY P,REDDY P.In vitro methyl farnesoate secretion by mandibular organs isolated from different molt and reproductive stages of the crab Oziotelphusa senex senex[J].Fisheries Science,2006,72(2):410-414.
• [22]NAGARAJU G,REDDY P,REDDY P.Mandibular organ:It's relation to body weight,sex,molt and reproduction in the crab,Oziotelphusa senex senex Fabricius(1791)[J].Aquaculture,2004,232:603-612.
• [23]李琛,郝瑞娟,王庆恒,等.马氏珠母贝磺基转移酶Pm CHST11基因的分子特征与表达分析[J].基因组学与应用生物学,2017,36(1):150-157.
• [24]傅郁.海湾扇贝(Argopecten irradians)外套膜组织学组织化学研究[D].青岛:中国海洋大学,2007.
• [25]孙虎山,王宜艳,王平,等.栉孔扇贝外套膜和鳃粘液细胞的类型与分布[J].中国水产科学,2002,9(4):315-317.
• [26]刘雅,郑哲,王庆恒,等.马氏珠母贝Pm-CTSC基因的克隆与表达分析[J].基因组学与应用生物学,2018,37(11):4723-4729.
• [27]曹艳飞,曾森林,焦钰,等.马氏珠母贝(Pinctada fucata martensii)细胞粘附分子3(Pm-CAM3)基因的克隆与表达分析[J].基因组学与应用生物学,2017.[2017-09-25].http://kns.cnki.net/kcms/detail/45.1369.Q.20170924.1531.010.html
• [28]吴羽媛,郭志颖,梁海鹰,等.马氏珠母贝Tlr6基因的克隆、序列分析与表达[J].水产学报,2017,41(11):1687-1698.
• [29]雷倩楠,崔少峰,梁海鹰,等.马氏珠母贝TRAF7基因c DNA的克隆及表达分析[J].基因组学与应用生物学,2016,35(1):79-86.
• [30]焦钰,杜晓东,王庆恒,等.马氏珠母贝IKKε同源基因的克隆及表达分析[J].基因组学与应用生物学,2014,33(2):266-272.
• [31]WU Y Y,LIANG H Y,WANG Z X,et al.A novel toll-like receptor from the pearl oyster Pinctada fucata martensii is induced in response to stress[J].Comparative Biochemistry and Physiology B:Biochemistry&Molecular Biology,2017,214:19-26.
• [32]孟勐,程道军,彭健,等.家蚕法尼酸甲基转移酶(FAMe T)基因的鉴定及表达模式分析[J].蚕业科学,2013,39(4):680-688.
• [33]杜俊俏,刁晓平,郑鹏飞,等.芘暴露对马氏珠母贝鳃和肝胰腺抗氧化酶活性的影响[J].生态环境学报,2013,22(10):1711-1716.
• [34]王志新,梁海鹰,杜晓东,等.马氏珠母贝热休克蛋白Hsp60基因的克隆与表达分析[J].广东海洋大学学报,2013,33(6):14-23.
• [35]李嘉尧.红螯光壳螯虾(Cherax Quadricarinatus)卵黄发生的研究[D].上海:华东师范大学,2008.
• [36]李志敏,李健,李吉涛,等.脊尾白虾(Exopalaemon carinicauda)FAMeT基因在卵巢发育周期中的表达分析[J].渔业科学进展,2016,37(1):46-51.