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1.
板栗CmAPs基因的克隆及在芽变短雄花序中的表达分析 总被引：1，自引：0，他引：1
利用RT-PCR与RACE扩增方法,分别从板栗正常雄花序和芽变短雄花序cDNA中分离出编码板栗天冬氨酸蛋白酶cDNA全长的序列,序列分析表明:克隆的cDNA片段总长分别为1 822 bp和1 909 bp,基因内部含有完整的开放阅读框架,大小均为1 539 bp,可编码长度为513个氨基酸残基的蛋白质,所推导的蛋白质氨基酸序列与蓖麻、可可和葡萄的天冬氨酸蛋白酶的蛋白质氨基酸序列相似性分别为83%,80%和75%.正常与芽变短雄花序中天冬氨酸蛋白酶的氨基酸序列存在6个氨基酸的差异,其中有3个位于活性区.后证实分离出的2个天冬氨酸蛋白酶基因在正常雄花序和芽变短雄花序中共同存在,命名为CmAPs1和CmAPs2(GenBank登录号分别为GQ984143和GQ084144).实时荧光定量PCR结果显示:在雄花序原基形成期、花簇原基形成期和花朵原基形成期,芽变花序中的CmAPs的总表达量均高于正常花序中的总表达量,而在发育的花被原基形成期即程序性死亡发生期,CmAPs在芽变花序中表达量远远低于正常花序中表达量.初步推测CmAPs与短雄花序发育过程中的程序性死亡有关. 相似文献
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以油茶近成熟种子为材料,根据油茶EST文库中已知的脂氢过氧化物裂解酶EST序列,利用3’RACE与5’RACE技术,克隆到脂氢过氧化物裂解酶的全长cDNA序列。该基因全长1648bp,,包含一个1476bp开放阅读框长,编码491个氨基酸,5’与3’非编码分别为52bp、121bp。预测该蛋白相对分子量为54.7779KDa,等电点为6.77,是个叶绿体转运肽,N端包含有22个疏水氨基酸残基组成的序列,不仅具有转运肽富含的丝氨酸,还含有大量转运肽中很少存在的脯氨酸。多序列比对发现油茶脂氢过氧化物裂解酶核苷酸序列与茶的相似性达到96%,因此将该基因命名coHPL。 相似文献
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天冬氨酸蛋白酶在天女木兰种子萌发过程中发挥着重要作用。以天女木兰种子为试验材料,利用同源克隆法RACE技术,从天女木兰种子中克隆得到天冬氨酸蛋白酶基因,全长1 545 bp,可以编码514个氨基酸残基。同源性分析显示,目的序列推导的氨基酸序列与其它植物中已知的天冬氨酸蛋白酶基因的氨基酸序列的同源性都在85%以上;采用生物信息学的方法进行分析,初步预测该基因编码的蛋白属于跨膜的分泌蛋白,大部分区域属于疏水结构;结构域的预测分析显示,此类蛋白包含约100个植物组织蛋白酶所特有的区域,该区域可能在植物液泡中发挥着重要作用。本研究为下一步系统、全面地研究天女木兰种子休眠的分子机理奠定了基础。 相似文献
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丙酮酸激酶是糖酵解途径的关键酶,在油料作物脂肪酸合成中有重要作用。在实验室构建的油茶EST文库基础上,采用5′RACE和交错延伸PCR技术获得了油茶PK基因的全长cDNA克隆。该基因序列长2 114 bp,开放读码框为1 737 bp,编码579个氨基酸,蛋白分子量为63 766.5 Da,分子式为C2774H4470N784O875S30,与其他植物的PK基因具有较高的相似性。PK基因系统进化树表明,油茶与蓖麻、杨树、葡萄的亲缘关系最近。 相似文献
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为进一步探索臣龙竹速生与巨大的分子机理,研究其AINTEGUMENTA（ANT）基因功能,研究试验以巨龙竹cDNA第一链为模板,用简并引物、基因特异引物以及运用RACE技术首次克隆出巨龙竹ANT基因全长cDNA序列。其序列全长为1914bp,与水稻、玉米、小麦等植物同类基因序列相似性高达85％;并且含有一个837bp的开放阅读框,编码278个与水稻、拟南芥和烟草ANT蛋白氨基酸序列有着较高同源性的巨龙竹ANT前体蛋白氨基酸序列。 相似文献
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以宇佐美曲霉E001总RNA为模板,采用RT-PCR等技术扩增了木聚糖酶基因(xynI)的cDNA全序列(GenBank登录号DQ302412)。该cDNA全长881 bp,其中5′和3′端非编码区分别为97和106 bp,信号肽编码区111 bp,成熟肽编码区567 bp编码188个氨基酸组成的木聚糖酶Xyn I。以E001基因组DNA为模板,采用单侧PCR等技术扩增了xynI的DNA全序列(GenBank登录号DQ302413)。该DNA全长1 098 bp,含启动子序列、内含子和外显子等核苷酸序列。将xynI cDNA所推导的Xyn I一级结构与GenBank中报道的其它相关序列进行了同源性比较。结果表明,E001 Xyn I是一种新的木聚糖酶,且具有第G/11家族木聚糖酶的共同特征。 相似文献
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以宇佐美曲霉E001总RNA为模板,采用RT- PCR等技术扩增了木聚糖酶基因(xyn I)的cDNA全序列(GenBank登录号DQ302412).该cDNA全长 881 bp,其中5′和3′端非编码区分别为97和 106 bp,信号肽编码区 111 bp,成熟肽编码区 567 bp 编码188个氨基酸组成的木聚糖酶Xyn I.以E001基因组DNA为模板,采用单侧PCR等技术扩增了xyn I的DNA全序列(GenBank登录号DQ302413).该DNA全长 1 098 bp,含启动子序列、内含子和外显子等核苷酸序列.将xyn I cDNA所推导的Xyn I一级结构与GenBank中报道的其它相关序列进行了同源性比较.结果表明,E001 Xyn I是一种新的木聚糖酶,且具有第G/11家族木聚糖酶的共同特征. 相似文献
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《中南林业科技大学学报(自然科学版)》2015,(7)
桐油是制备生物柴油的优势原料。delta 8脱饱和酶基因与植物膜脂、鞘磷脂的生物合成及细胞信号传递途径有关。通过简并PCR扩增、3’RACE、5’RACE及编码区扩增获得了油桐delta 8脱饱和酶的全长c DNA序列。该基因全长1 787 bp,ORF的长度为1 344 bp,编码447个氨基酸。经过网上比对分析,发现油桐delta 8脱饱和酶与其它植物的此基因具有较高同源性,其中与蓖麻脂肪酸去饱和酶相似度达83%。研究结果为揭示油桐油脂合成途径及分子定向育种奠定基础。 相似文献
10.
以杜仲叶片cDNA为模板,采用反转录RCR及RACE技术分离出HDR基因的cDNA克隆,命名为EuHDR。EuHDR基因cDNA全长1 653 bp,5’端非编码区长82 bp,3’端非编码区长188 bp,编码460个氨基酸,与喜树HDR基因序列相似性最高,达82%;推导EuHDR氨基酸序列中包含转运肽序列(A1-A33)及植物HDR蛋白多个保守的功能位点(A117,A208,A262,A345);EuHDR蛋白二级结构α-螺旋占35.65%,β-折叠占19.78%,螺环结构占44.57%;EuHDR蛋白三级结构为单体形式,呈不规则的三叶草形状;系统进化分析表明EuHDR蛋白与葡萄HDR蛋白的亲缘关系最为接近。 相似文献
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In this paper, the basic principle of chromosome walking is presented and we used an actin gene of radiata pine (Pinus radiata) as an example to conduct upstream and downstream chromosome walking for EST sequences. The full genomic sequence (2154 bp)
of the actin gene, including promoters 5′ UTR, CDS and 3′ UTR, was identified by chromosome walking. PCR amplification and DNA band sequencing
from 200 unrelated radiata pine trees revealed a total of 21 SNPs for the actin gene, three in the promoter region, 15 in CDS and 4 in 3′ UTR. The results of this experiment provide a technical framework
for SNPs discovery in none coding regions of candidate genes.
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Translated from Acta Botanica Boreali-Occidentalia Sinica, 2007, 27 (8): 1571–1576 [译自: 西北植物学报] 相似文献
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Yuzhi Dong Chuanping Yang Daoyuan Zhang Yucheng Wang 《Frontiers of Forestry in China》2007,2(2):217-221
Plant aquaporins are water-selected-channels in plants and are involved in seed germination, cell elongation, stoma movement,
fertilization and so on. Some plant aquaporins also play an important role in drought stress response. In this paper, the
gene encoding the Tamarix albiflonum Aquaporin (AQP) was amplified by 5′rapid amplification of cDNA end (RACE) on the basis of the sequence information obtained
from the expressed sequence tag of the subtractive hybridization library constructed under PEG6000 stress. The cDNA of the
T. albiflonum AQP gene is 1,043 bp long, encoding a protein of 287 amino acids with a predicted molecular mass of 30.9 kDa, has 6 transmembrane
regions, and possessing the major intrinsic protein (MIP) family signal consensus sequence SGXHXNPAVT and the higher plant
plasma membrane intrinsic protein (PIP) highly conservative sequence GGGANXXXXGY and TGI/TNPARSL /FGAA I/VI/VF/YN. A comparative
molecular analysis of the nucleotide sequence in National Center for Biotechnology Information (NCBI) databases showed that
it shared 95% homology with the gene of Arabidopsis thaliana (MIP-C), with a theoretical isoelectric point 8.84.
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Translated from Bulletin of Botanical Research, 2006, 26(4): 475–479 [译自: 植物研究] 相似文献
15.
在转录组测序结果分析基础上,以山鸡椒cDNA 为模板,克隆得到山鸡椒1-脱氧木酮糖-5-磷酸还原异构酶DXR 基因cDNA 全长,以山鸡椒基因组DNA 为模板,设计引物、扩增拼接后获得山鸡椒DXR 基因全长,命名为LcDXR。序列分析表明,LcDXR cDNA 全长为1 501 bp,5'非编码区长34 bp,3'非编码区长53 bp,开放阅读框长1 413 bp,预测编码含有470个氨基酸残基的蛋白质,等电点为6.62,分子量为51.12 kD。LcDXR 基因全长为12 601 bp,其中外显子12 个,内含子11 个。对来自10个种源的LcDXR 基因编码区单核苷酸变异位点进行分析表明:在cDNA 区间内共发现10 个SNP(single nucleotide polymorphism)位点,其中有4 个单核苷酸变异导致了所编码的氨基酸的改变,为了分析氨基酸突变导致的蛋白质精细结构的变化,利用Swiss-PDB Viewer 模拟4 个突变位点氨基酸残基的替换。其中江西安远(AY)的突变Lys119Thr 引起了氢键的变化,推测可能对酶的活性产生影响。研究结果为深入研究山鸡椒脱氧木酮糖5-磷酸还原异构酶的活性和功能奠定了基础,同时为山鸡椒遗传育种提供理论依据。 相似文献
16.
YANG Qian YU Han-ying LI Chang-yin 《林业研究》2005,16(4):260-264
An atom gene was cloned from genomic DNA of Scleortinia sclerotiorum by inverse PCR. The evolutionary relationships of S. sclerotiorum and other fungi in atom gene were studied. Results showed that the atom gene from of S. sclerotiorum has a single open reading frame of 4 773 bp and does not include any introns. The derived amino acid sequence consists of 1 590 residues, and it is homologous to all fungal AROM proteins studied so far. The theoretical isoelectric point （pl） and molecular weight （Mw） is 6.5 and 172.66 kD, respectively GC percentage of the arom gene is 44.94. According to the results of searching from CDD and Prosite database, AROM protein of S. sclerotiorum contains five conserve domains： 3-dehydroquinate synthase domain, 3-dehydroquinate dehydratase （3-dehydroquinase） domain, shikimate 5-dehydrogenase domain, shikimate kinase domain, and -enolpyruvylshikimate-3-phosphate synthase （EPSP sythase） domain, and four motifs： two EPSP synthase signatures, dehydroquinase class I active site, shikimate kinase signature. According to the PIR Site Rule PIRSR000514-1, four functionally important amino acid residues are found by alignment. Putative TATA box and CAAT box locate separately in -23 and -77 loci in 5＇ un-translated region, and two loci found in downstream atom gene are likely polyadenylation signals. In addition, phylogeny of atom gene is analyzed. 相似文献
17.
Construction and analysis of a subtracted cDNA library of Betula platyphylla female inflorescence 总被引：2，自引：0，他引：2
WEIJi-cheng YANGChuan-ping WANGChao JIANGJing 《林业研究》2005,16(2):97-100
Female inflorescence of Betula platyphylla was sampled at an interval of each two days to analyze the background of gene expression in floral phase. On the basis of SMART strategy, the driver cDNA was obtained from total RNA of the last sample and the tester cDNA was from that of the others by RT-PCR which were subsequently used to construct a subtracted cDNA library. The result of the ESTs (expression sequence tags) blastX showed that the genes in the subtracted cDNA library could be mainly clustered into 5 groups related to metabolism, transportation and signal transduction, cell cycle, stress response, and regulation. The relationship between gene expression and development was also discussed. 相似文献
18.
APETALA2(AP2)基因在植物生长发育过程中发挥着重要作用.利用RT-PCR和RACE方法,从毛竹中克隆到1个AP2同源基因的全长cDNA序列,命名为PeAP2.序列分析表明:PeAP2基因全长1 750 bp,其中,5′端非编码区106bp,3′端非编码区174 bp,开放阅读框1 470 bp,编码1个489 aa的蛋白,该蛋白含有2个AP2结构域,属于AP2/EREBP家族的AP2亚家族.PeAP2蛋白与来自其它单子叶植物的AP2蛋白均有着较高同源性,其中,与二穗短柄草的AP2蛋白同源性最高,达74.85％.实时定量PCR分析显示:PeAP2基因在毛竹的根、茎、叶、鞘和节5种器官中均有表达,其中,叶片中的表达丰度最高,鞘中次之,而在根、茎、节中的表达丰度接近,均较低.利用hiTAIL-PCR方法克隆获得了PeAP2上游启动子区序列1 359 bp,分析显示其含有光、激素等多种信号应答相关的作用元件. 相似文献
19.
Naoko Yamaguchi Susumu Shiraishi Yoshinori Sasaki Chiaki Yamamoto 《Journal of Forest Research》2000,5(1):35-38
Leyland cypress (×Cupressocyparis leylandii) has been regarded as an intergeneric hybrid between Monterey cypress (Cupressus macrocarpa) and Alaska cypress (Chamaecyparis nootkatensis) in the Cupressaceae. Each partial sequence of the 18S-rDNA from nuclear and therbcL from chloroplast DNA genomes was determined using above three species. At the 59th site from the 5′ end of the 152 bp of
the determined 18S-rDNA sequence, one base difference was identified between Monterey cypress (T) and Alaska cypress (A). However, Leyland cypress
showed either nucleotide A or T at this site. Since nuclear DNA is inherited by biparental mode, this result shows genetic
evidence that the nucleic genome of Leyland cypress originates from Monterey cypress and Alaska cypress and that this species
is of hybrid origin. 相似文献