犬弓首蛔虫abcg-5基因的克隆及序列分析
|
摘要
研究犬弓首蛔虫(Toxocara canis,T.canis)腺苷三磷酸结合盒转运蛋白(ATP-binding cassette transporters, ABC)亚家族G5基因(abcg-5)的分子特征.根据犬弓首蛔虫基因组数据库中的Tc-abcg-5基因序列设计引物,通过PCR技术克隆Tc-abcg-5全长基因,并进行多重序列比对和种系发育进化树分析.结果表明:该基因全长为1 902 bp,编码633个氨基酸.功能结构域分析发现TcABCG5蛋白包含1个ABC转运蛋白结构域和6个跨膜区,同时发现了高度保守的Walker A和Walker B模体. GO分析显示ABCG5具有ATP结合和ATP酶活性.种系发育进化树分析显示TcABCG5与猪蛔虫进化关系较近,与哺乳动物进化关系较远.
To study the molecular characteristics of ATP-binding cassette sub-family G member 5(abcg-5) in Toxocara canis, the full-length coding sequence of Tc-abcg-5 gene was amplified using PCR amplification, multiple sequence alignment(MSA) was made and a phylogenetic tree was constructed. The results showed that the entire sequence of Tc-abcg-5 gene contained 1 902 bp, encoding a protein of 633 amino acids. TcABCG5 was found to contain one ABC transporter domain and six transmembrane regions. The highly conserved Walker A and Walker B motifs of the ABCG sub-family were also found. Gene ontology(GO) annotations showed that TcABCG5 had the molecular function of ATP binding and ATPase activity. Phylogenetic analysis indicated that TcABCG5 shared a high level of amino-acid sequence similarity with Ascaris suum, and shared a low level of amino-acid sequence similarity with mammals. This study is expected to lay a foundation for further functional researches of TcABCG5.
To study the molecular characteristics of ATP-binding cassette sub-family G member 5(abcg-5) in Toxocara canis, the full-length coding sequence of Tc-abcg-5 gene was amplified using PCR amplification, multiple sequence alignment(MSA) was made and a phylogenetic tree was constructed. The results showed that the entire sequence of Tc-abcg-5 gene contained 1 902 bp, encoding a protein of 633 amino acids. TcABCG5 was found to contain one ABC transporter domain and six transmembrane regions. The highly conserved Walker A and Walker B motifs of the ABCG sub-family were also found. Gene ontology(GO) annotations showed that TcABCG5 had the molecular function of ATP binding and ATPase activity. Phylogenetic analysis indicated that TcABCG5 shared a high level of amino-acid sequence similarity with Ascaris suum, and shared a low level of amino-acid sequence similarity with mammals. This study is expected to lay a foundation for further functional researches of TcABCG5.
引文
[1]OVERGAAUW P A,VAN ZUTPHEN L,HOEK D,et al.Zoonotic Parasites in Fecal Samples and Fur from Dogs and Cats in The Netherlands[J].Veterinary Parasitology,2009,163(1/2):115-122.
[2]MACPHERSON C N.The Epidemiology and Public Health Importance of Toxocariasis:a Zoonosis of Global Importance[J].International Journal for Parasitology,2013,43(12/13):999-1008.
[3]SCHMITT I,TAMPE R.Structure and Mechanism of ABC Transporters[J].Current Opinion in Structural Biology,2002,16(2):754-760.
[4]GREENBERG R M.Schistosome ABC Multidrug Transporters:from Pharmacology to Physiology[J].International Journal for Parasitology,Drugs and Drug Resistance,2014,4(3):301-309.
[5]SEYFFER F,TAMPE R.ABC Transporters in Adaptive Immunity[J].Biochimica et Biophysica Acta,2015,1850(3):449-460.
[6]BLOISE E,ORTIGA-CARVALHO T M,REIS F M,et al.ATP-Binding Cassette Transporters in Reproduction:a New Frontier[J].Human Reproduction Update,2016,22(2):164-181.
[7]JONES P M,GEORGE A M.Multidrug Resistance in Parasites:ABC Transporters,P-Glycoproteins and Molecular Modelling[J].International Journal for Parasitology,2005,35(5):555-566.
[8]ARDELLI B F.Transport Proteins of the ABC Systems Superfamily and their Role in Drug Action and Resistance in Nematodes[J].Parasitology International,2013,62(6):639-646.
[9]吴春太,李维国,高新生,等.植物DNA甲基转移酶的生物信息学分析[J].西南大学学报(自然科学版),2010,32(4):83-89.
[10]HOWE K L,BOLT B J,SHAFIE M,et al.WormBase ParaSite-A Comprehensive Resource for Helminth Genomics[J].Molecular and Biochemical Parasitology,2017,215(7)::2-10.
[11]HOPFNER K P.Invited Review:Architectures and Mechanisms of ATP Binding Cassette Proteins[J].Biopolymers,2016,105(8):492-504.
[12]JONES P M,GEORGE A M.The ABC Transporter Structure and Mechanism:Perspectives on Recent Research[J].Cellular and Molecular Life Sciences:CMLS,2004,61(6):682-699.
[13]CONSEIL G,DEELEY R G,COLE S P.Functional Importance of Three Basic Residues Clustered at the Cytosolic Interface of Transmembrane Helix 15in the Multidrug and Organic Anion Transporter MRP1(ABCC1)[J].The Journal of Biological Chemistry,2006,281(1):43-50.
[14]CHOI Y H,YU A M.ABC Transporters in Multidrug Resistance and Pharmacokinetics,and Strategies for Drug Development[J].Current Pharmaceutical Design,2014,20(5):793-807.
[15]HONORAT M,FALSON P,TERREUX R,et al.Multidrug Resistance ABC Transporter Structure Predictions by Homology Modeling Approaches[J].Current Drug Metabolism,2011,12(3):268-277.
[16]WENZEL J J,PIEHLER A,KAMINSKI W E.ABC A-Subclass Proteins:Gatekeepers of Cellular Phospho and Sphingolipid Transport[J].Frontiers in Bioscience:a Journal and Virtual Library,2007,12(5):3177-3193.
[17]SCHULZ B,KOLUKISAOGLU H U.Genomics of Plant ABC Transporters:the Alphabet of Photosynthetic Life forms or Just Holes in Membranes[J].FEBS Letters,2006,580(4):1010-1016.
[18]PATRAWALA L,CALHOUNal T,SCHNEIDER-BROUSSARD R,et al.Side Population is Enriched in Tumorigenic,Stem-Like Cancer Cells,Whereas ABCG2+and ABCG2-Cancer Cells Are Similarly Tumorigenic[J].Cancer Research,2005,65(14):6207-6219.
[19]VERRIER P J,BIRD D,BURLA B,et al.Plant ABC Proteins-a Unified Nomenclature and Updated Inventory[J].Trends in Plant Science,2008,13(4):151-159.
[20]TRAN P N,BROWN S H,MITCHELL T W,et al.A Female Gametocyte-Specific ABC Transporter Plays a Role in Lipid Metabolism in the Malaria Parasite[J].Nature Communications,2014,5(9):4773-5773.
[21]KASINATHAN R S,MORGAN W M,GREENBERG R M.Genetic Knockdown and Pharmacological Inhibition of Parasite Multidrug Resistance Transporters Disrupts Egg Production in Schistosoma mansoni[J].PLoS Neglected Tropical Diseases,2011,5(12):e1425.
[2]MACPHERSON C N.The Epidemiology and Public Health Importance of Toxocariasis:a Zoonosis of Global Importance[J].International Journal for Parasitology,2013,43(12/13):999-1008.
[3]SCHMITT I,TAMPE R.Structure and Mechanism of ABC Transporters[J].Current Opinion in Structural Biology,2002,16(2):754-760.
[4]GREENBERG R M.Schistosome ABC Multidrug Transporters:from Pharmacology to Physiology[J].International Journal for Parasitology,Drugs and Drug Resistance,2014,4(3):301-309.
[5]SEYFFER F,TAMPE R.ABC Transporters in Adaptive Immunity[J].Biochimica et Biophysica Acta,2015,1850(3):449-460.
[6]BLOISE E,ORTIGA-CARVALHO T M,REIS F M,et al.ATP-Binding Cassette Transporters in Reproduction:a New Frontier[J].Human Reproduction Update,2016,22(2):164-181.
[7]JONES P M,GEORGE A M.Multidrug Resistance in Parasites:ABC Transporters,P-Glycoproteins and Molecular Modelling[J].International Journal for Parasitology,2005,35(5):555-566.
[8]ARDELLI B F.Transport Proteins of the ABC Systems Superfamily and their Role in Drug Action and Resistance in Nematodes[J].Parasitology International,2013,62(6):639-646.
[9]吴春太,李维国,高新生,等.植物DNA甲基转移酶的生物信息学分析[J].西南大学学报(自然科学版),2010,32(4):83-89.
[10]HOWE K L,BOLT B J,SHAFIE M,et al.WormBase ParaSite-A Comprehensive Resource for Helminth Genomics[J].Molecular and Biochemical Parasitology,2017,215(7)::2-10.
[11]HOPFNER K P.Invited Review:Architectures and Mechanisms of ATP Binding Cassette Proteins[J].Biopolymers,2016,105(8):492-504.
[12]JONES P M,GEORGE A M.The ABC Transporter Structure and Mechanism:Perspectives on Recent Research[J].Cellular and Molecular Life Sciences:CMLS,2004,61(6):682-699.
[13]CONSEIL G,DEELEY R G,COLE S P.Functional Importance of Three Basic Residues Clustered at the Cytosolic Interface of Transmembrane Helix 15in the Multidrug and Organic Anion Transporter MRP1(ABCC1)[J].The Journal of Biological Chemistry,2006,281(1):43-50.
[14]CHOI Y H,YU A M.ABC Transporters in Multidrug Resistance and Pharmacokinetics,and Strategies for Drug Development[J].Current Pharmaceutical Design,2014,20(5):793-807.
[15]HONORAT M,FALSON P,TERREUX R,et al.Multidrug Resistance ABC Transporter Structure Predictions by Homology Modeling Approaches[J].Current Drug Metabolism,2011,12(3):268-277.
[16]WENZEL J J,PIEHLER A,KAMINSKI W E.ABC A-Subclass Proteins:Gatekeepers of Cellular Phospho and Sphingolipid Transport[J].Frontiers in Bioscience:a Journal and Virtual Library,2007,12(5):3177-3193.
[17]SCHULZ B,KOLUKISAOGLU H U.Genomics of Plant ABC Transporters:the Alphabet of Photosynthetic Life forms or Just Holes in Membranes[J].FEBS Letters,2006,580(4):1010-1016.
[18]PATRAWALA L,CALHOUNal T,SCHNEIDER-BROUSSARD R,et al.Side Population is Enriched in Tumorigenic,Stem-Like Cancer Cells,Whereas ABCG2+and ABCG2-Cancer Cells Are Similarly Tumorigenic[J].Cancer Research,2005,65(14):6207-6219.
[19]VERRIER P J,BIRD D,BURLA B,et al.Plant ABC Proteins-a Unified Nomenclature and Updated Inventory[J].Trends in Plant Science,2008,13(4):151-159.
[20]TRAN P N,BROWN S H,MITCHELL T W,et al.A Female Gametocyte-Specific ABC Transporter Plays a Role in Lipid Metabolism in the Malaria Parasite[J].Nature Communications,2014,5(9):4773-5773.
[21]KASINATHAN R S,MORGAN W M,GREENBERG R M.Genetic Knockdown and Pharmacological Inhibition of Parasite Multidrug Resistance Transporters Disrupts Egg Production in Schistosoma mansoni[J].PLoS Neglected Tropical Diseases,2011,5(12):e1425.