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砂梨ACC合酶cDNA全长克隆及其反义与正义表达载体构建

DOI: 10.7685/j.issn.1000-2030.2008.04.005, PP. 25-31

Keywords: 砂梨,ACC合酶,cDNA,反义/正义表达载体

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Abstract:

为构建干扰砂梨(PyruspyrifoliaNakai)ACC合酶基因表达的遗传转化载体,利用RACE技术从’早生新水’成熟果实cDNA中克隆了ACC合酶的cDNA,该cDNA全长为1939bp,命名为Pyp-ACS,GenBank登录号为EF566865。Pyp-ACS核苷酸序列含有5′末端非翻译区109bp、1488bp完整的开放阅读框和3′末端非翻译区342bp(含25bp的Ploy+(A))。Pyp-ACS编码495个氨基酸,与白梨(Pyrus×bretschneideriRehd.)、西洋梨(PyruscommunisL.)、中国李(PrunussalicinaLindl.)、桃(Prunuspersica(L.)Batsch)、梅(PrunusmumeSeib.etZucc.)、苹果(Malus×domesticaBorkh.)和柿(DiospyroskakiThunb.)有较高的同源性。该氨基酸序列具备ACC合酶7个保守区和组成该酶活性中心的12个氨基酸残基,即SLSKDMGFPGLR,进一步验证了克隆的正确性。以pYPX145载体为基础,分别将Pyp-ACS编码区序列反向和正向插入相应位点构建反义和正义表达载体,并分别命名为pPyp-ACS(-)和pPyp-ACS(+),目的基因由双35S启动子所控制。分别将这2个表达载体导入根癌农杆菌菌株(Agrobacteriumtumefaciens)EHA105,为耐贮藏转基因梨的遗传转化提供载体。

References

[1]  Oeller P W,Lu M W,Taylor L P.Reversible inhibition of tomato fruit senescence by antisense RNA[J].Science.1991,254:437-439
[2]  Dandekar A M,Teo G,Defilippi B G.Effect of down-regulation of ethylene biosynthesis on fruit flavor complex in apple fruit[J].Transgenic Research.2004,13:373-384
[3]  Gao M,Matsuta N,Murayama H.Gene expression and ethylene production in transgenic pear(Pyrus communis cv.’La France’)with sense or antisense cDNA encoding ACC oxidase[J].Plant Science.2007,173:32-42
[4]  吴静,李瑞珍,徐碧玉.香蕉ACC合成酶反义基因转化香蕉的研究[J].分子植物育种.2007,5(4):497-501
[5]  Manning K.Isolation of nucleic acids from plants by differential solvent precipitation[J].Analytical Biochemistry.1991,195(1):45-50
[6]  黄骥,张红生.基于Windows的核酸序列分析软件的开发[J].生物信息学.2004,2(1):13-17
[7]  Dower W J,Miller J F,Ragsdale C W.High efficiency transformation of E.coli by high voltage electroporation[J].Nucleic Acids Research.1988,16(13):6127-6145
[8]  Sambrook J,Russell D W.Molecular Cloning:A Laboratory Manual[M].New York:cold Spring Harbor Laboratory Press.2001
[9]  Rottmann W H,Peter G F,Oeller P W.1-aminicyclopropane-1-carboxylate synthase in tomato is encoded by a multigene family whose transcription is induced during fruit and floral senescence[J].Journal of Molecular Biology.1991,22:937-961
[10]  Hayama H,Shimada T,Fujii H.Ethylene-regulation of fruit softening and softening-related genes in peach[J].Journal of Experimental Botany.2006,57:4071-4077
[11]  Shiomi S,Yamamoto M,Ono T.cDNA cloning of ACC synthase and ACC oxidase in genes in cucumber fruit and their differential expression by wounding and auxin[J].Japanese Society for Horticulture Science.1998,67:685-692
[12]  El-Sharkawy I,Lelièvre J M,Pech J C.ACC synthase genes related to cold-dependent ripening in pear fruit[J].Acta Horticulturae.2005,682:355-362
[13]  刘翠琼,汤浩茹.梨叶片培养与转基因研究进展[J].果树学报.2003,20(5):374-378
[14]  Dong J G,Kim W T,Yip W K.Cloning of a cDNA encoding 1-aminocyclopropane-1-carboxylate synthase and expression of its mRNA in ripening apple fruit[J].Planta.1991,185:38-45
[15]  Sunako T,Ishikawa R,Senda M.MdACS-5A(accession No.AB034992)and 5B(accession No.AB034993),two wound-responsive genes encoding 1-aminocyclopropane-1-carboxylate synthase in apple[J].Plant Physiology.2000,122:620
[16]  Ikoma Y,Yano M,Ogawa K.Cloning and expression of genes encoding ACC synthase in kiwifruit[J].Acta Horticulturae.1995,398:179-186
[17]  Whittaker D J,Smith G S,Gardner R C.Expression of ethylene biosynthetic genes in Actinidia chinensis fruit[J].Plant Molecular Biology.1997,34:45-55
[18]  Mathooko F M,Tsunashima Y,Owino W Z O.Regulation of genes encoding ethylene biosynthetic enzymes in peach(Prunus persica L.)fruit by carbon dioxide and 1-methylcyclopropene[J].Postharvest Biology and Technology.2001,21:265-281
[19]  饶景萍,童斌,中野龙平.柿果实ACC合成酶cDNA的克隆及其序列分析[J].西北植物学报.2001,21(5):819-825
[20]  van der Straeten D,van Wiemeersch L,Goodman H M.Cloning and sequence of two different cDNAs encoding 1-aminicyclopropane-1-carboxylate synthase in tomato[J].Proceedings of the National Academy of Sciences(USA).1990,87:4859-4863
[21]  Yip W K,Dong J G,Kenny J W.Characterization and sequencing of the active-site of 1-aminicyclopropane-1-carboxylate synthase[J].Proceedings of the National Academy of Sciences(USA).1990,87:7930-7934
[22]  Kaneyoshi J,Wabiko H,Kobayashi S.Agrobacterium tumefaciens AKE10-mediated transformation of an Asian pear,Pyrus betulaefolia Bunge:host specificity of bacterial strains[J].Plant Cell Reports.2001,20:622-628
[23]  周莉莉,蔡斌华,乔玉山.不同处理对丰水梨离体叶片不定芽再生的影响[J].南京农业大学学报.2007,30(2):34-38
[24]  Hood E E,Gelvin S B,Melchers L S.New Agrobacterium helper plasmids for gene transfer to plants[J].Transgenic Research.1993,2:208-218
[25]  Kondo S,Isuzugawa K,Kobayashi S.Aroma volatile emission and expression of 1-aminocyclopropane-1-carboxylate(ACC)synthase and ACC oxidase genes in pears treated with 2,4-DP[J].Postharvest Biology and Technology.2006,41:22-31

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