首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 62 毫秒
1.
花生△12-脂肪酸脱氢酶基因的克隆及序列分析   总被引:2,自引:1,他引:1  
根据已报道的△12-脂肪酸脱氢酶基因(FAD)的氨基酸保守序列设计引物进行RT-PCR扩增,得到花生△12-脂肪酸脱氢酶基因881bp部分cDNA序列,然后通过快速扩增cDNA末端技术(RACE),向两端延伸得到1410bp的花生△12-脂肪酸脱氢酶基因全长cDNA序列。序列分析表明有一个长1140bp、编码379个氨基酸残基的开放阅读框,所编码蛋白质的大小约为43kDa。推测的氨基酸序列具有膜整合蛋白酶特异性的3个组氨酸保守区;氨基酸疏水性分析结果表明,所编码的氨基酸序列存在2个具有膜固定蛋白(membrance-anchored protein)重要特征的疏水结构,2个疏水区共跨膜4次,这些特性表明所获得的序列为△12-脂肪酸脱氢酶基因。  相似文献   

2.
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   

3.
以一编码蛋白与拟南芥的CAB1高度相似的EST序列作为探针,通过同源搜索从橡胶树的转录组数据中电子克隆到一条长1 415 bp的cDNA,该序列包含一798 bp的ORF及47 bp的5′UTR和570 bp的3′UTR,命名为HbCAB1;在此基础上,采用PCR技术成功扩增了包括该ORF在内的915 bp的cDNA序列。序列分析表明,该基因预测编码265个氨基酸,理论分子量为28.15 ku,等电点为5.25;蛋白含有1个保守的捕光叶绿素a/b结合蛋白结构域,3个跨膜螺旋和1个C端螺旋,1个三聚化基序,  相似文献   

4.
参考龙眼转录组数据库信息,以莆田宝庵寺的龙眼古树“宝树”幼胚诱导的胚性愈伤组织作为试验材料,获得了龙眼古树wrky44基因的全长序列,命名为Dlwrky44,登录号为JF709012,共2313 bp,其中3′UTR458 bp,poly A尾长25 bp,5′UTR 735 bp,并包含1个1119 bp的开放阅读框,编码373个氨基酸.根据推导的氨基酸序列进行生物信息学分析结果表明:D1WRKY44蛋白是不稳定、亲水蛋白;无信号肽和跨膜结构,亚细胞定位于细胞核中;具有2个WRKY结构域,并具有核定位信号,在wrky家族中属第1类.系统进化树分析结果表明:龙眼古树DlWRKY44蛋白与柽柳亲缘关系最近.对龙眼古树EC Dlwrky44基因在不同低温胁迫条件下(0、5、10、15、20、25℃)处理12h的表达进行了分析,结果表明:Dlwrky44基因的表达量随着温度的逐渐降低,表达量逐渐升高,当温度升到15℃时,表达量达到最高.  相似文献   

5.
以负调控禾本科植物分蘖的关键基因TB1为研究对象,根据水稻、玉米、高粱TB1同源基因保守序列设计引物,使用RT-PCR方法和RACE技术从甘蔗茎尖处克隆到该基因的同源基因ScTB1。ScTB1 cDNA全长1 668 bp,包含274 bp的5′ UTR,1 149 bp的CDS和245 bp的3′ UTR。生物信息学分析表明该基因可编码382个氨基酸残基,编码产物含有保守的SP区、TCP区和R区,属于TCP家族转录因子。其分子量为40.7 ku,理论等电点为8.55,蛋白亚细胞定位预测其主要定位于细胞质。系统进化分析表明ScTB1属于CYC/TB1亚族蛋白,与高粱TB1和玉米TB1聚为一个亚类。根据序列的保守性和预测结果可推测ScTB1很可能也参与了甘蔗分蘖性状的调控。  相似文献   

6.
孙化鹏  钟晓红  乔飞 《热带作物学报》2018,39(11):2200-2206
甲羟戊酸焦磷酸脱羧酶(Mevalonate diphosphate decarboxylase,MVD)是三萜皂苷生物合成途径中的关键酶, 为研究其在洋常春藤中的基因功能,通过 RT-PCR 和 RACE 技术从洋常春藤叶片中克隆获得 HhMVD 基因的 cDNA 全 长序列,并通过 RT-qPCR 技术分析其表达规律。结果表明:RACE 克隆获得的 HhMVD 基因 cDNA 序列全长 1 799 bp, 包含一个完整开放阅读框(ORF)1 263 bp、5′非编码区(5′UTR)192 bp、3′非编码区(3′UTR)344 bp;该基因编码 420 个氨基酸,分子质量 46.6 ku,理论等电点为 6.57,不含跨膜区,属于非分泌型蛋白;HhMVD 蛋白具有 GHMP 激 酶 N 端结构域,属于甲羟戊酸激酶系列,与刺五加、人参、三七等同科植物亲缘关系较近。RT-qPCR 结果表明,洋常 春藤中 HhMVD 基因的时空表达相对稳定,但与常春藤皂苷含量差异变化之间存在一定的负相关性。洋常春藤 HhMVD 基因的成功克隆及表达分析研究,为深入探讨其基因功能、阐明其在常春藤皂苷生物合成途径中的作用提供理论依据。  相似文献   

7.
利用橡胶树胶乳EST文库克隆到一个TUA基因,命名为HbTUA1(GenBank登录号:KC333454)。该基因cDNA全长1 580 bp,其中5′UTR长45 bp,3′UTR长167 bp,编码区长1 368 bp,编码449个氨基酸。HbTUA1蛋白具有TUA类蛋白保守的GTP结合域。荧光定量PCR分析显示,HbTUA1基因在橡胶树胶乳中的表达量最高,其次是树皮和芽,而在种子和叶片中的表达量最低;在胶乳中,HbTUA1基因的表达显著受割胶和伤害诱导,在不同死皮程度的橡胶树中也存在明显差异。初步表明HbTUA1基因可能参与橡胶树胶乳再生与胁迫应答调控。  相似文献   

8.
以铁皮石斛原球茎为材料,采用同源克隆的方法,成功得到了铁皮石斛Do-HDR基因的全长,并通过q PCR对其相对表达量进行分析。结果表明:Do-HDR基因全长1 784 bp(Gen Bank登录号KJ946381),5′UTR为40 bp,3′UTR为361 bp,开放阅读框为1 382 bp,编码460个氨基酸。生物信息学分析结果表明,该基因编码的蛋白为稳定的亲水蛋白,不具有跨膜结构,不含信号肽,定位于内质网。采用灭活的尖孢镰刀菌菌液作为诱导子,分别以0、100、200、500、1 000 mg/L的浓度培养3 d后测定总生物碱含量,结果表明,总生物碱含量呈现先上升后下降的趋势;当诱导子浓度为200 mg/L时,总生物碱含量达到最大值。在此基础上通过q PCR分析Do-HDR基因的相对表达量,结果发现Do-HDR基因相对表达量在诱导子浓度为200 mg/L时也到达最大值。因此,推测Do-HDR基因与生物碱的合成与积累有关。  相似文献   

9.
γ-ECS基因编码的γ-谷氨酰半胱氨酸合成酶是细胞内谷胱甘肽(GSH)从头合成的限速酶。以龙眼转录组数据库为基础,采用同源克隆的方法从龙眼胚性愈伤组织中分离得到γ-ECS基因cDNA全长序列,在GenBank上的登录号为JF815477。γ-ECS基因序列全长1 812 bp,包括17 bp 5′UTR及254 bp 3′UTR;ORF长度为1 541 bp编码511个氨基酸。生物信息学分析结果表明,龙眼胚性愈伤组织γ-ECS的核甘酸序列及其推导的氨基酸序列分别与蓖麻、毛果杨等具有较高的同源性;γ-ECS为1个具有跨膜结构的非分泌蛋白,具有亲水性;亚细胞定位于过氧化物酶体,不含信号肽,并存在亮氨酸拉链结构。龙眼胚性愈伤组织γ-ECS基因在龙眼体胚发生过程中的整体表达趋势是先升高,后降低,其表达量在心形胚阶段达到最大值之后在鱼雷形胚阶段急剧下降。γ-ECS基因表达量变化可能与GSH在龙眼体胚发生过程中的含量变化一致。  相似文献   

10.
为进一步了解小分子量热激蛋白在小麦耐热能力中的作用,根据玉米16.9 kD小分子热激蛋白的氨基酸序列,采用同源序列法进行序列拼接和引物设计,用RT-PCR扩增获得了1个源自小麦叶片的热激蛋白基因cDNA片段,即TaHSP16.9-1(GenBank登录号为EU649679).TaHSP16.9-1全长770 bp,5′非翻译区81 bp,3′非翻译区233 bp,开放阅读框编码151个氨基酸.序列分析结果表明,此蛋白与已知单子叶植物来源的同类基因高度同源,相似性介于78.3%~96.7%之间.定量RT-PCR表达谱分析显示,TaHSP16.9-1在小麦抽穗期的茎和旗叶以及幼苗期叶片中均能表达,其在小麦幼苗叶片中的表达受高温胁迫的诱导.  相似文献   

11.
A collection of 173 Triticum tauschii accessions was analysed to evaluate the variability of low molecular weight (Mr) glutenin subunits. These proteins were analysed by one-step one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and were divided into B-, C- and D-subunits in accordance with their electrophoretic mobility. Extensive polymorphism, both in the number and electrophoretic mobility, was detected in lowMr glutenin subunits present in T. tauschii. Thirty different patterns for B-subunits and forty-three for C-subunits were identified, some of which were with identical electrophoretic mobility than those observed in hexaploid wheat. Glutenin subunits with the same electrophoretic mobilities of low Mr D-glutenin subunits as well as subunits encoded at the Glu-D4 and Glu-D5 loci, were also detected in accessions of T. tauschii. These results provide new basic knowledge regarding the genetics variability of the low Mr glutenin subunits, as well as their potential to create novel germplasm for the improvement of wheat quality in breeding programs.  相似文献   

12.
An enzyme-linked immunosorbent assay (ELISA) was used to determine the bifunctional alpha-amylase/subtilisin inhibitor (BASI) content of barley grain from 11 cultivars grown in six diverse locations in Australia. The inhibitor ranged from 119 to 254 μg/g in 57 barley samples. Genotype had a significant (P<0·05) effect on BASI content but there was no effect due to environment. Total protein varied independently of BASI and was influenced by environment and genotype. BASI content was higher (P<0·05) in malting barley than in feed barley and was correlated positively (r=0·29;P<0·05) with alpha-amylase activity in corresponding malts. The ELISA used monoclonal and polyclonal antibodies raised against purified BASI. In immunoblot analysis the monoclonal antibody showed high specificity for the inhibitor in barley and also detected the inhibitor in wheat. Low levels of inhibitor (mean 3·2 μg/g) were found in 12 Australian wheat cultivars using the ELISA developed for barley. The assay had a linear working range of 5–50 ng/mL with a detection limit of 2 ng/mL. Reproducibility between assays was good (CV=4·9%) but mean recoveries were high, ranging from 116–129% when purified inhibitor was added to barley extracts. The ELISA may have useful applications in brewing research and barley breeding programmes.  相似文献   

13.
杂交育种是巴西橡胶树品种遗传改良的重要途径。以近年来在中国引种试种成功的国外胶木兼优无性系为亲本之一进行人工杂交授粉工作,研究其杂交子代苗期的性状表现,为胶木兼优品种的创新利用提供参考依据。分析热垦509×热研87-4-26、热研78-2×热垦628和热研78-2×热垦523等3个杂交组合子代苗期生长、试割干胶产量、乳管分化能力等性状。结果表明:组合内各性状存在较大差异,不同杂交组合子代试割前年茎围增粗均大于8 cm,试割后年均茎围增粗4.5 cm以上,均优于对照RRIM600;组合热垦509×热研87-4-26子代的综合性状均表现较好,亲本的速生和高产性状能较好的遗传给后代,从该组合子代中选育出胶木兼优品种的可能性较大。  相似文献   

14.
Hemicellulose acetates were prepared under homogeneous reaction conditions in the system N,N-dimethylacetamide/lithium chloride by reacting the native hemicelluloses with acetic anhydride in the presence of 4-dimethylaminopyridine within 72 h at 60–85°C. The products obtained were characterised by means of Fourier transform infrared chromatography, gel permeation chromatography, and thermal analysis. The degree of substitution of acetylated hemicelluloses ranged between 0.74 and 1.49 as a function of experimental conditions. Under an optimum reaction condition (85°C, 60 h), over 80% hydroxyl groups in native hemicelluloses were acetylated. The molecular weight measurements showed that a significant degradation and hydrolysis of the products appeared at only a prolonging period of 72 h. It was found that the thermal stability of the products increased by esterification.  相似文献   

15.
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   

16.
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   

17.
金龟子绿僵菌对红棕象甲的室内致病力测定   总被引:1,自引:0,他引:1  
在室内用喷雾法测定13株绿僵菌对红棕象甲的致病力。结果表明,菌株致病力强弱与菌株种类有密切关系,菌株的浓度与累计死亡率成线性关系。菌株对红棕象甲的致病力由强到弱的顺序为:Red2>Red3>Gre1>Yx7>Yx4>Gre3>Red1>Yx1>Gre2>Yx6>Gre4>Gre5>Yx3。接种Red2菌株在孢子浓度为1.0×108个/mL水悬浮剂情况下,对4龄红棕象甲幼虫的LT50为5.23 d。用不同浓度的Red2菌株的孢子油悬浮剂涂抹法接种4龄幼虫,发现其LC50为7.32×106个/mL。另外,经试  相似文献   

18.
Helitron是一类新发现的DNA转座元件,具有捕获宿主基因片段的能力,因而对植物基因及基因组的进化具有重要意义。为了研究麦类作物中这类元件对基因组的进化所起的作用,本研究采用一种改进的局部组合变量(LCV)方法对大麦基因组中的Helitron转座元件进行了预测,共鉴定了16 560个潜在的Helitron元件,去除假阳性后这些元件约占基因组的2.2%。基于序列分歧的计算发现,绝大多数Helitron形成于最近的900万年内,并且在大约250和550万年前各经历了一次剧烈扩张。在大麦基因组中发现了3个自主型Helitron元件,它们包含Rep和Helicase结构域的编码区。基因片段捕获分析发现,66.5%的元件平均捕获了1.9个基因片段,除了反转座和转座相关的基因外,参与发育调控的NAM家族转录因子和参与水分运输和环境胁迫响应的软木脂素合成酶基因也被高频捕获,表明Helitron转座元件不仅影响大麦基因组的结构,同时可以驱动特定功能基因的扩增。  相似文献   

19.
稻瘟病菌(Magnaporthe oryzae)是研究病原真菌生长发育及致病机理的重要模式生物。Rho家族蛋白是一类具有GTP酶活性的GTP结合蛋白,在多种细胞信号转导通路中起着分子开关的作用。通过生物信息学方法分析Mo Rho2的理化性质、蛋白结构域等信息,并推测其潜在功能。为进一步明确Mo Rho2蛋白的功能,利用基因敲除技术获得Mo Rho2基因的敲除突变体。表型分析结果表明,与野生型菌株相比,Mo Rho2敲除突变体的菌丝生长速度、分生孢子形态、附着胞形态、产孢量、对植物的毒力等方面均无明显的差异,但分生孢子萌发和附着胞形成略滞后于野生型菌株。突变体能够产生正常的侵入栓并侵入洋葱表皮。综上所述,Mo Rho2基因可能参与稻瘟病菌分生孢子的萌发和附着胞的形成,结果为进一步揭示基因Mo Rho2的生物学功能奠定基础。  相似文献   

20.
为研究巴西橡胶树钾转蛋白(KT/HAK/KUP)基因家族在生长发育和逆境胁迫方面的生物学功能,从橡胶树基因组中鉴定得到31个KT/HAK/KUP基因家族成员,并从基因结构、染色体定位、系统进化和表达分析等方面进行全面系统的分析。研究结果发现,KT/HAK/KUP基因家族31个成员分布在3个Contig和15个染色体上,8号染色体上数目较多为8个成员,其中5个为串联重复;各成员编码的氨基酸残基数目在288~881个不等,蛋白分子量分布在31.82~98.96 kDa,其蛋白产物均定位于细胞质膜上;内含子数目为5~11个,进化上分为4个明显分支。在表达方面,部分成员呈现明显的组织特异性,在叶片发育过程中成员HbKUP5HbKUP6HbKUP9HbKUP14表达发生明显变化,乙烯利刺激后HbKUP30HbKUP31在胶乳中的表达明显上调,在抗寒品种‘93~114’中,HbKUP5HbKUP9在低温处理后明显下调表达。本研究结果为深入解析KT/HAK/KUP基因家族的功能和培育高钾抗逆的橡胶树品种提供了理论基础。  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号