胡钟东;乔玉山;陶建敏;渠慎春;章镇;

• 1:南京农业大学园艺学院
• 2:南京农业大学园艺学院
• 3:南京农业大学园艺学院
• 4:南京农业大学园艺学院

摘要(Abstract)：

根据已报道的砂梨ACO3(GenBank Accession No.AB042107)序列设计并合成一对特异引物,以砂梨品种若光(Wakahikari)叶片总DNA为模板,利用染色体步移技术扩增基因的上游调控序列,回收该片段并克隆到pMD19-T载体,测序后去除接头和基因编码序列。得到1段长715bp的上游序列,登陆PLACE数据库在线进行植物DNA顺式作用元件预测并分析,起始位点的上游89bp处有1个TATA-Box核心启动子元件。另外序列还存在多个特异调控基因表达的元件,初步判断为砂梨果实中ACC氧化酶基因的诱导型启动子。

关键词(KeyWords)： 砂梨;ACC氧化酶;调控序列;序列分析

Abstract:

Keywords:

基金项目(Foundation): 江苏省高技术研究(BG2006313);; 南京农业大学青年基金(KJ05007)

作者(Author): 胡钟东;乔玉山;陶建敏;渠慎春;章镇;

Email:

DOI:

参考文献(References)：

• [1]LUJ,ZHAOH Y,HE YK,SONGYR.Advances and progresses of high promoter research and its application[J].Natural Science Pro-gresses,2004,14(8):856-862.(in Chinese)路静,赵华燕,何奕昆,宋艳茹.高等植物启动子及其应用研究进展[J].自然科学进展,2004,14(8):856-862.
• [2]ZAREMBINSKI T I,THEOLOGIS A.Ethylene biosynthesis and ac-tion:a case of conservation[J].Plant Mol Biol,1994,26:1579-1597.
• [3]ROTTMANN W H,PETER G F,OELLER P W,KELLER J A,SHENNF,NAGYBP,TAYLOR LP,CAMPBELLAD,THEOL-OGIS A.1-aminocyclopropane-l-carboxylate synthase in tomato is encoded by a multi-gene family whose transcription is induced dur-ingfruit and floral senescence[J].J Mol Biol,1991,222:937-961.
• [4]BARRY C S,BLUME B,BOUZAYEN M,COOPER W,HAMIL-TON A J,GRIERSON D.Differential expression of the1-aminocy-clopropane-1-carboxylate oxidase gene family of tomato[J].Plant J,1996,9:525-535.
• [5]TANG X Y,WANG H,BRANDT A S,WOODSON W R.Organiza-tion and structure of the1-aminocyclopropane-1-carboxylate oxi-dase gene family fromPetunia hybrida[J].Plant Mol Biol,1993,23:1151-1164.
• [6]KIMW T,YANG S F.Structure and expression of cDNAs encoding1-aminocyclopropane-1-carboxylate oxidase homologs isolated from excised mungbean hypocotyls[J].Planta,1994,194(2):223-229.
• [7]ITAI A,KAWATA T,TANABE K,TAMURA F,UCHIYAMA M,TOMOMITSU M,SHIRAIWA N.Identification of1-aminocyclo-propane-1-carboxylic acid synthase genes controlling the ethylene level ofripeningfruit in Japanese pear(Pyrus pyrifolia Nakai)[J].Mol Gen Genet,1999,261(1):42-49.
• [8]LOPEZ G R,CAMPBELL A,DONG J G,YANG S F,GOMEZ-LIM M A.Ethylene biosynthesis in banana fruit:isolation of a ge-nomic clone to ACC oxidase and expression studies[J].Plant Sci,1997,123:123-131.
• [9]WANGXL,PENGXX.Cloningofpromoter ofbanana fruit ripening related ACO1and primary study on its function[J].Chinese Journal ofBiotechnology,2001,17(4):428-431.(in Chinese)王新力,彭学贤.香蕉果实成熟相关基因ACO1启动子区的克隆及其功能初探[J].生物工程学报,2001,17(4):428-431.
• [10]MOON H,CALLAHAN A M.Developmental regulation of peach ACC oxidase promoter-GUS fusions in transgenic tomato fruits[J].J Exp Bot,2004,55(402):1519-1528.
• [11]TAI T H,TANKSEY D D.Rapid isolation of total DNA from dehy-drated plant tissue[J].Plant Mol Biol Rep,1990,8(4):297-303.
• [12]PRIBNOW D.Nucleotide sequence of an RNA polymerase binding site at an earlyT7promoter[J].Proc Natl Acad Sci,1975,72:784-788.
• [13]BREATHNACH R,CHAMBON P.Organization and expression of eucaryotic split genes coding for proteins[J].Annu Rev Biochem,1981,50:349-383.
• [14]SINGER V L,WOBBLE C R,STRUHL K.A wide variety of DNA sequences can functionally replace a yeast TATA element for tran-scriptional activation[J].Genes&Dev,1990,4:636-645.
• [15]LAGRANGE T,KAPANIDIS A N,TANG H,REINBERG D,E-BRIGHT R H.New core promoter element in RNA polymerase II-dependent transcription:Sequence-specific DNA binding by tran-scription factor IIB[J].Genes&Dev,1998,12:34-44.
• [16]SHIRSATA,WILFORDN,CROY R,BOULTER D.Sequences re-sponsible for the tissue specific promoter activity of a pea legumin gene in tobacco[J].Mol Gen Genet,1989,215:326-331.
• [17]FUJIWARAT,BEACHYR N.Tissue-specific and temporal regula-tion ofa beta-conglycinin gene:roles ofthe RYrepeat and other cis-actingelements[J].Plant Mol Biol,1994,24:261-272.
• [18]NAG R,MAITY MK,DASGUPTA M.Dual DNA binding property of ABA insensitive3like factors targeted to promoters responsive to ABAand auxin[J].Plant Mol Biol,2005,59:821-838.
• [19]RIEPING M,SCHOFFL F.Synergistic effect of upstream sequences CCAAT box elements,and HSE sequences for enhanced expression of chimaeric heat shock genes in transgenic tobacco[J].Mol Gen Genet,1992,231(2):226-232.
• [20]SCHARFKD,ROSES,ZOTTW,SCH#FFLF,NOVERL,SCH#FF F.Three tomato genes code for heat stress transcription factors with domain ofthe yeast HSF[J].EMBOJ,1900,9:4495-4501.
• [21]GAGLIARDI D,BRETON C,CHABOUD A,VERGNE P,DUMAS C.Expression of heat shock factor and heat shock protein70genes duringmaize pollen development[J].Plant Mol Biol,1995,29:841-856.
• [22]HUBEL A,SCHOFFL F.Arabidopsis heat shock factor:Isolation and characterization of the gene and the recombinant protein[J].Plant Mol Biol,1994,26:352-362.
• [23]SARGE KD,PARKKS,KIRBY J D,MAYO K E,MORIMOTO R I.Expression of heat shock factor2in mouse testis:potential role asa regulator of heat shock protein gene expression during spermatoge-nesis[J].Biol Reprod,1994,50:1334-1343.
• [24]KIMS Y,CHUNG H J,THOMAS T L.Isolation of a novel class of bZIP transcription factors that interact with ABAresponsive and em-bryo specification elements in the Dc3promoter using a modified yeast one-hybrid system[J].Plant J,1997,11:1237-1251.
• [25]FINKELSTEIN R R,LYNCH T J.The Arabidopsis abscisic acid re-sponse gene ABI5encodes a basic leucine zipper transcription factor[J].Plant Cell,2000,12:599-609.
• [26]LOPEZ-MOLINA L,CHUA N H.A null mutation in a bZIP factor confers ABA-insensitivity in Arabidopsis thaliana[J].Plant Cell Physiol,2000,41:541-547.
• [27]EZCURRA I,ELLERSTROM M,WYCLIFFE P,STALBERG K,RASK L.Interaction between composite elements in the napA pro-moter:both the B-boxABA-responsive complexand the RY/Gcom-plex are necessary for seed specific expression[J].Plant Mol Biol,1999,40(4):699-709.
• [28]EZCURRA I,WYCLIFFE P,NEHLIN L,ELLERSTROM M,RASK L.Transactivation of the Brassica napus napin promoter by ABI3re-quires interaction of the conserved B2and B3domains of ABI3with different cis-elements:B2mediates activation through an ABRE,whereas B3interacts with an RY/G-box[J].Plant J,2000,24(1):57-66.
• [29]HO S N,HUNT H D,HORTON R M,PULLEN J K,PEASE L R.Site-directed mutagenesis byoverlap extension using the polymerase chain reaction[J].Gene,1989,77(1):51-59.