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1.
利用染色体步移的方法克隆了一种白木香倍半萜合酶——愈创木烯合酶基因(AsSGS)791 bp的启动子序列,序列分析表明,获得的序列中A/T含量达64.6%,与真核生物启动子序列的特征相符合。该序列除具有启动子核心序列TATA-box和增强子元件CAAT-box等典型的真核生物启动子基本元件外,还含有如赤霉素反应元件GARE-motif、ABA的应答元件G-Box、生长素响应元件TGA-element和脱落酸响应元件ABRE等参与激素调控元件,光应答元件Box I、GA-motif、TCT-motif,热胁迫反应的HSE,厌氧诱导元件ARE和增强诱发厌氧反应的GC-motif等顺式作用元件。利用农杆菌介导的本生烟草瞬时表达系统对所获得的启动子功能进行鉴定,结果表明,所有缺失片段都能驱动β-葡萄糖苷酸酶(GUS)基因表达,GUS活性与启动子片段的长度呈正相关。 相似文献
2.
茶树CsANS基因及其启动子的克隆与生物信息学分析 总被引：1，自引：0，他引：1
采用RACE技术,获得茶树武夷奇种C18叶片花青素合成途径中的花青素合成酶(ANS)基因的c DNA全长序列。采用染色体步移技术获得该基因的启动子序列。采用实时荧光定量PCR技术检测该基因在不同遮光处理下的表达动态。结果表明,Cs ANS全长c DNA为1 000 bp,其中ORF(Open Reading Frame)为957 bp,编码320个氨基酸,含有DIOX-N和20G-Fell-Oxy保守功能结构域;分离得到Cs ANS基因上游调控序列1 010bp,其含启动子核心元件TATA-box及ACE、GT1-motif、Sp1(光响应元件)、circadian(生物钟相关元件)等与花青素合成途径相关的重要顺式作用元件。荧光定量PCR分析表明,该基因在全光照处理(CK)时表达量较高,75%遮光时表达量较低。说明该基因的表达受光照强弱的控制。 相似文献
3.
利用染色体步移法克隆了白木香查尔酮合成酶基因(As CHS1)ATG上游1 082 bp的启动子序列,该启动子序列中AT含量高达69.03%,符合真核生物启动子的序列特征。通过启动子预测软件分析可知,该序列的转录起始位点位于翻译起始位点-65 bp处,并且其上游-25~30 bp区域存在TATA-box等典型的真核生物启动子核心元件,同时含有一些顺式作用元件如赤霉素应答元件GARE-motif、茉莉酸甲酯应答元件CGTCA-motif、水杨酸应答元件TCA-element等激素调控元件,光应答元件BoxⅠ、G-Box、ACE,厌氧诱导元件ARE等。通过构建p C-1 082pro As CHS1植物表达载体,借助农杆菌将重组载体转化到烟草叶片中。蛋白定量结果表明,该序列可以驱动GUS的表达,具有启动子活性;脱落酸显著增强该启动子的活性,乙烯则显著抑制该启动子的活性。 相似文献
4.
利用生物信息学方法分析陆地棉脱水素基因GhDHN1的DNA序列结构特性,预测其功能。序列分析表明,GhDHN1基因的基因组DNA中AT碱基含量为52.16%,而内含子具有较高的AT碱基含量（63.33%）;GhDHN1基因不含CpG岛;其内含子序列含有核心启动子元件TATA框、启动子的增强子元件CAAT框,并含有参与光响应的保守DNA模块的部分元件（Box 4）、赤霉素响应元件GARE-motif、生长素响应元件TGA元件等,说明该内含子可能参与了该基因的转录活动,可能受光、激素的诱导;GhDHN1内含子具有GT-AG 规则的保守序列,推测GhDHN1基因具有保守的内含子剪接机制。 相似文献
5.
分离克隆龙眼(Dimocarpus longan Lour.)胚性愈伤组织14-3-3基因,并分析该基因在龙眼体胚发生过程中的表达情况。采用RT-PCR结合RACE法,获得龙眼胚性愈伤组织14-3-3基因的cDNA全长序列和DNA序列,采用TAIL-PCR法,获得龙眼胚性愈伤组织14-3-3基因的启动子DNA序列,运用生物信息学方法对序列进行分析,并通过实时荧光定量PCR(qRT-PCR)研究该基因在龙眼体胚发生过程中的表达。结果表明:经克隆得到龙眼胚性愈伤组织14-3-3基因786 bp的cDNA全长序列(GenBank检索号为GU573765),该cDNA开放阅读框推定的氨基酸序列(含261个氨基酸)与其它植物14-3-3具有较高同源性;该基因的DNA序列(GenBank检索号为GU573766)长为1 859 bp,包含3个内含子,内含子的剪切位点均符合真核生物"GT-AG"规则;该基因的启动子DNA序列(GenBank检索号为GU573766)长为910 bp;该基因在龙眼体胚各阶段均有表达,其中在松散型胚性愈伤组织阶段和心形胚及鱼雷形胚阶段呈现高表达,整个变化趋势呈"倒S"状。 相似文献
6.
为探究夜来香（Cestrum nocturnum）花香基因SAMT在花香代谢中的调控作用,以花瓣为材料,采用RT-PCR和RACE技术相结合,克隆夜来香SAMT基因的全长cDNA。结果表明：该cDNA的序列长度为1 493 bp,编码365个氨基酸,其中包含1 098 bp ORF、130 bp 5′UTR和265 bp 3′UTR,将其命名为CnSAMT;生物信息学分析结果表明：该基因所编码的氨基酸序列与烟草、番茄的同源性分别为98％和98％,属于甲基转移酶-7家族;原核表达结果表明：CnSAMT的蛋白表达产物约为42 ku,与软件预测的结果大致相同。采用染色体步移技术,克隆夜来香CnSAMT的5′端调控序列,结果表明：该序列长度为993 bp,且该序列除了含有TATA-box、CAAT-box等启动子核心元件外,还含有光、生长素、脱落酸、热胁迫、缺氧胁迫等外界环境条件响应的顺式作用元件。 相似文献
7.
以茶树新品系“1005”嫩芽为材料,克隆得到CsMYB基因gDNA全长序列1β828βbp,通过染色体步移技术,分离得到CsMYB基因启动子片段1β038βbp,命名为proMYB。生物信息学分析表明,CsMYB gDNA由2个内含子和3个外显子组成,该启动子含有与提高基因转录水平相关的5′UTR Py-rich stretch顺式作用元件、启动子核心元件TATA-box和CAAT-box,还存在有激素响应元件、光响应元件、逆境应答元件以及大量功能未知或功能特异的顺式作用元件,说明proMYB具有诱导型启动子特性,CsMYB基因在茶树植物体中可能参与多种非生物胁迫应答和激素信号传导;通过烟草的瞬时表达结果表明,该启动子能驱动下游基因表达,具有启动子活性。 相似文献
8.
为深入研究巴西橡胶树HbWRKY1基因的表达调控机制,采用染色体步移法克隆了HbWRKY1基因的5′调控区序列。结果表明,该序列具有预测的核心启动子区域位于-6~-57 bp,转录起始位点位于翻译起始位点上游41 bp处;该序列具有TATA-box,CAAT-box和GATA-box等多个典型的真核生物启动子基本的顺式作用元件,同时还具有低温响应元件LTR,赤霉素反应元件GARE-motif,生长素响应元件TGA-element和脱落酸响应元件ABRE等参与激素调控元件,此外还有与MYB结合的MBS元件,热胁迫反应的HSE,厌氧诱导元件ARE和增强诱发厌氧反应的GC-motif等顺式作用元件。构建了不同长度的HbWRKY1启动子片段的植物表达载体并转化本生烟草, 通过潮霉素抗性和PCR鉴定,获得了转基因植株。对获得的转HbWRKY1不同长度启动子片段的转基因T1植株GUS染色发现,植株的茎、叶柄、主叶脉和根中均可呈现蓝色,表明HbWRKY1基因的5′调控区可以启动GUS基因的表达, 具有启动子的活性。 相似文献
9.
以甘蔗品种FN28为材料,首次克隆甘蔗ShSUT4基因。测序结果表明,该基因约为4.8 kb,包含5个外显子和4个内含子,并包括完整的开放阅读框。该基因5＇侧翼序列长度约为1.8 kb。采用PLACE、PlantCARE在线启动子预测工具分析表明,该序列含有启动子的特定结构,如TATA-box、CAAT-box等,还含有一些顺式作用元件如光响应元件、MYB结合位点、ABRE响应元件等,表明甘蔗ShSUT4基因的表达可能受光照、MYB和ABRE等的调控。 相似文献
10.
为深入研究巴西橡胶树HbWRKY1基因的表达调控机制,采用染色体步移法克隆了HbWRKY1基因的5'调控区序列.结果表明,该序列具有预测的核心启动子区域位于-6～-57 bp,转录起始位点位于翻译起始位点上游41 bp处;该序列具有TATA-box,CAAT-box和GATA-box等多个典型的真核生物启动子基本的顺式作用元件,同时还具有低温响应元件LTR,赤霉素反应元件GARE-motif’生长素响应元件TGA-element和脱落酸响应元件ABRE等参与激素调控元件,此外还有与MYB结合的MBS元件,热胁迫反应的HSE,厌氧诱导元件ARE和增强诱发厌氧反应的GC-motif等顺式作用元件.构建了不同长度的HbWRKY1启动子片段的植物表达载体并转化本生烟草,通过潮霉素抗性和PCR鉴定,获得了转基因植株.对获得的转HbWRKY1不同长度启动子片段的转基因T1植株GUS染色发现,植株的茎、叶柄、主叶脉和根中均可呈现蓝色,表明HbWRKY1基因的5'调控区可以启动GUS基因的表达,具有启动子的活性. 相似文献
11.
In Vitro 《Journal of Cereal Science》1996,24(3):215-225
Resistant starch (RS), producedin vitroby hydrolysis of retrograded pea starch gels and amylose gels by porcine pancreaticalpha-amylase, was characterised by X-ray diffraction, size exclusion chromatography and methylation analysis. These techniques showed that RSin vitroconsisted of semi-crystalline, mostly linear material that was present in two main molecular size subfractions (DPn>100 andDPn20–30) with a third, minor subfraction (DPn≤5). The extent of retrogradation of amylose was found to be of primary importance in determining the RS content of starch. Analysis ofin vivoRS, recovered during an ileostomy study, produced results that were similar to those obtained from RSin vitro. Anin vitromodel for the structure of resistant starch is proposed. 相似文献
12.
P1B-type ATPase ion pumps that transport heavy metal ions across cellular membranes are essential for plant growth and development. To date, a genomic comparison overview of the family in rice, maize and sorghum is not yet available. In this study, a total of 31 heavy metal P1B-type ATPase (HMA) genes were identified, including 9 in rice, 11 each from maize and sorghum. They were classified into two distinct subfamilies based on their sequence composition and phylogenetic relationship. Four pairs of HMA genes were expanded via gene duplication with tandemly duplicated. Comprehensive analyses were performed to investigate the expression profiles of HMA genes in various tissues by using quantitative real-time PCR. Some HMA members exhibited abundant and tissue-specific expression patterns. Moreover, most of the genes were found to be differentially expressed under the Cu/Cd treatment. This study will facilitate further studies on P1B-type ATPase family and provide valuable hints for the functional validation in rice, maize and sorghum. 相似文献
13.
根据一个从巴西橡树胶乳cDNA文库中获得的EST片段的序列设计引物,通过RACE的方法获得了橡胶树编码annexin的cDNA,命名为AnnHb1。序列分析表明,AnnHb1长为1 198 bp,含有945 bp的阅读框,62 bp的5'-UTR和191 bp的3'-UTR,编码314个氨基酸,分子量为35.99 ku,等电点为8.18。该氨基酸序列与蓖麻、麻疯树、棉花、苜蓿、烟草和拟南芥中的annexin的同源性分别为82%、72%、72%、64%、60%和60%。半定量RT-PCR分析结果表明AnnHb1基因在愈伤、花、树皮、叶、胶乳中均有表达,其中在愈伤组织中表达量最低,树皮中表达量最高。 相似文献
14.
Carboxyl group-terminated poly(N-isopropylacrylamide) (PNIA-COOH) was synthesized via radical polymerization of N-isopropylacrylamide (NIA) using mercaptoacetic acid (MAA) as a chain transfer agent. The molecular weight of the PNIA-COOH
was controlled by changing the molar ratio of MAA to NIA. A water-soluble chitosan derivative, N-(2-hydroxy)propyl-3-trimethylammonium chitosan chloride (HTCC), was also synthesized by reacting chitosan with glycidyltrimethylammonium
chloride. Then, chitosan-g-PNIA and HTCC-g-PNIA copolymers were synthesized using the “graft-onto” method by reacting PNIA-COOH with chitosan and HTCC, respectively.
The formation of the grafted copolymers was confirmed by Fourier transform infrared spectroscopy, solubility test in water,
and scanning electron microscopy — energy dispersive spectroscopy. The thermo-responsive behaviors of the grafted copolymers
and the change in lower critical solution temperature (LCST) were also studied. Chitosan-g-PNIA was insoluble in water and behaved like a thermo-responsive hydrogel due to the crosslinking-point action of the chitosan
backbone. The swelling ratio of chitosan-g-PNIA increased with increasing PNIA content. HTCC-g-PNIA behaved as a water-soluble thermo-responsive polymer. Compared to the homo PNIA, the LCST of HTCC-g-PNIA was slightly increased. 相似文献
15.
Byoung Chul Chun Tae Keun Cho Mi Hwa Chong Yong-Chan Chung David Martin Jihua Chen Jong-Shin Park 《Fibers and Polymers》2007,8(1):43-49
Nanocomposite of polyurethane (PU), Nylon66 (nylon), and montmorillonite (MMT) was prepared by a twin screw extruder, and
the dispersion of MMT and the mechanical properties of the nanocomposite were analyzed. Dimethyl hydrogenated tallow 2-ethylhexyl
ammonium modified Cloisite 25A (C25A) and methyl tallow bis-2-hydroxyethyl ammonium modified Cloisite 30B (C30B) were used
as MMT. XRD and TEM analysis indicated that the continuous melt mixing by a twin screw extruder was effective in MMT dispersion.
C30B having hydroxyl group on its surface has better dispersion than C25A in the PU/nylon matrix. Maximum stress and strain
at break were the maximum at 1 wt% MMT regardless of matrix composition, and decreased at higher MMT content. Best MMT dispersion
was also observed at 1 wt% MMT for the entire matrix composition. Aggregation of MMT occurred at MMT content higher than 1
wt%. Nylon addition also induced the aggregation of MMT because of the high polarity of nylon surface. Dispersion of MMT was
very important in improving the mechanical properties of PU/nylon nanocomposite. 相似文献
16.
日本稻米烘干·储藏·加工·流通·消费中的品质管理及信息追溯 总被引：1，自引：0，他引：1
从稻米的烘干、储藏、碾米加工等方面阐述了日本的稻米品质管理与品质评价。介绍了日本大米的JAS法标识和大米信息追溯系统的结构。以能够进行烘干、砻谷、选别、储藏、碾米的自动货架系统为中心,构想稻米联合设施,对稻米烘干储藏、加工流通体系进行展望。 相似文献
17.
18.
The activities of endogenous (R-type) and exogenous acting (D-type) protein inhibitors ofalpha-amylase and the activities ofalpha- and total amylase were determined in milling fractions of rye. High D-type amylase inhibitor activities were detected in the embryo (255 IU/g) and in the endosperm fraction (64·9 IU/g), low inhibitor activities were found in the aleurone layer fraction (25·9 IU/g). The highest R-typealpha-amylase inhibitor activity was found in the aleurone layer fraction (32·6 IU/g), and the lowest value in the epidermis containing fraction (5·0 IU/g). The D- and R-typealpha-amylase inhibitor activities varied with growing conditions. D-type amylase inhibitor activities were found to be high in those samples which grew under drought conditions and low in samples cultivated under wet and cool weather. Higher R-typealpha-amylase inhibitor activities were found in rye genotypes cultivated under wet conditions and lower values under dry weather. There were small variations inalpha-amylase inhibitor activities between sprout-stable and sprout-sensitive rye genotypes. The D- and R-typealpha-amylase inhibitor activities of all varieties were stable during 72 h of germination. Similar soil conditions will therefore lead to differentialalpha-amylase inhibitor activities depending on weather conditions during growth. 相似文献
19.
This research evaluates the miscibility and performance of polypropylene (PP)/polybutylene succinate (PBS) and PP/polylactic acid (PLA) blend and natural-flour-filled, PP/PLA and PP/PBS blend bio-composites. The melting temperature (T m ) and glass transition temperature (T g ) of pure PP, PBS and PLA showed a single peak but differential scanning calorimetry (DSC) and dynamic mechanical thermal analysis (DMTA) presented two peaks for the T m and T g of the PP/PBS and PP/PLA blends. These results indicated that the PP/PBS and PP/PLA blend systems existed as immiscible blends. These results were also confirmed by the scanning electron microscopy (SEM) micrographs of the tensile fracture surface of the PP/PBS and PP/ PLA blends. At a PP/PBS and PP/PLA blend ratio of 70/30, the tensile and flexural strengths of bamboo flour (BF)- and wood flour (WF)-filled, PP/PBS and PP/PLA blend bio-composites were similar to those of BF- and WF-filled, PP and PBS bio-composites. In addition, these strengths of maleic anhydride-grafted PP (MAPP)- and acrylic acid-grafted PP (AAPP)-treated, BF- and WF-filled, PP/PBS and PP/PLA blend bio-composites were higher than those of non-treated bio-composites. 相似文献
20.
以卡特兰紫色花品种‘粉女郎’（Cattleya hybrid‘Pink Lady’）为材料,采用RT-PCR和RACE方法从花瓣中分离得到了一个查尔酮合酶（chalcone synthase, CHS）和一个二氢黄酮醇4-还原酶（dihydroflavonol 4-reductase, DFR）同源基因的cDNA全长,分别命名为ChCHS1和ChDFR1,GenBank登录号分别为KP171693和KP171694。序列分析结果发现：ChCHS1的cDNA全长1 508 bp ,编码394个氨基酸;ChDFR1基因的cDNA全长1 250 bp,编码350个氨基酸。同源性检索和生物信息学分析表明,这2个基因编码的蛋白都具有各自的功能位点和保守性特征多肽序列。氨基酸序列比对与系统进化树分析显示,ChCHS1、ChDFR1基因在兰科中与蕙兰、石斛兰关系最近,与百合科关系较近。相对实时荧光定量PCR结果表明,ChCHS1 和ChDFR1都在蕾期表达量最低,伴随花朵的开放,表达量逐渐上升,最终分别在花朵盛开期和接近开放时达到最高。 相似文献