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LI Yan-he ZHENG Fang-liang CHEN Hong-quan WANG Han-zhong WANG Liu-quan XU Di-ping 《中国农业科学(英文版)》2009,8(3):369-379
The full-length cDNA sequence of prophenoloxidase was obtained through RACE technology. The complete cDNA sequence is 3 721-bp long, containing an open reading frame （ORF） of 1 881 bp, a 154-bp 5′-untranslated region, and a 1 686- bp 3′-untranslated region with three potential functional poly（A） signals （AATAAA）. The molecular mass of the deduced amino acid sequence （627 aa） was 72.3 kDa with an estimatedpI of 5.88. It contained putative copper-binding sites （copper A： 131, 135, 167 and copper B： 301,305, 341）, and a tentative complement-like motif （GCGWPDHL）. Eight potential N-linked glycosylation sites were predicted to be present in P. clarkii prophenoloxidase. Similar to those in other arthropod prophenoloxidases reported so far, no signal peptide was detected in the crayfish prophenoloxidase. The phylogenetic trees confirmed that P. clarkii prophenoloxidase was most closely related to that of freshwater crayfish P. leniusculus and more closely related to other crustacean prophenoloxidases from shrimp, prawn, and lobster than to the insect prophenoloxidases. Besides, two putative introns were found in this sequence of genomic DNA. 相似文献
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SU Song-kun ZHNEG Huo-qing CHEN Sheng-lu ZHONG Bo-xiong Stefan Albert 《中国农业科学(英文版)》2005,4(9):707-713
By screening the worker （Apis cerana cerana） heads cDNA library using a fragment of the mrjp3 gene of Apis cerana as probe, 120 positive clones were obtained. The clone containing A. cerana cerana MRJP3 （AccMRJP3） cDNA was selected. Based on the sequencing of the inserts of the positive clone, a sequence of AccMRJP3 cDNA which is 1 887 bp long including a poly （A） tail was obtained. The AccMRJP3 cDNA encompassed an open-reading frame （ORF） with 1 779 bp encoding 593 amino acids. The un-translated regions （UTR） of the 5′ end and 3′end are 46 bp and 160 bp in length, respectively. Similar to AmMRJP3 and AdMRJP3, the putative AccMRJP3 also has a repetitive region. The comparison of the repetitive region of AccMRJP3, AmMRJP3 and AdMRJP3 shows some differences between them. 相似文献
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LI Guo-liang CHANG Hui ZHOU Ren-gang 《中国农业科学(英文版)》2007,6(9):1043-1050
A novel J-domain protein gene was cloned from wheat （Triticum aestivum L.） using RT-PCR technology and named as TaJ. The J-domain protein is defined by the presence of a J-domain. The cDNA of T. aestivum gene, TaJ （GenBank accession number： DQ789026）, was 1263 bp and contained a complete open reading frame （ORF） encoding a J-domain protein of 420 amino acid residues. The predicted amino acid sequence of TaJ possesses three functionally essential domains： the Nterminal J-domain which includes the highly conserved HPD tripeptide, an adjacent domain that is rich in glycine and phenylalanine residues （G/F） and a Cysteine-rich zinc-finger domain with four repeats of CxxCxGxG that is important for protein interactions. The C-terminal of TaJ was -CAQQ, a farnesylation motif. The full-length deduced amino acid sequence of TaJ is highly homologous to J-domain proteins from various plant species. Southern blot analysis indicated that a single copy of TaJ existed in wheat genome. The expression pattern of TaJ performed by real-time PCR demonstrated that heat shock （HS） at 37℃ induced the expression of TaJ rapidly and strongly, but the response of the TaJ gene to cold stress was much slower than that to HS. Tissue-specific expression analysis showed that the expression level of TaJ gene was much higher in leaves than that in roots. 相似文献
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CHEN Guo-yue YANG Shi-dong LI Wei-tao LI Wei WEI Yu-ming ZHENG You-liang 《中国农业科学(英文版)》2009,8(12):1440-1447
Grain texture in barley （Hordeum vulgare L.） is an important quality character. The single nucleotide polymorphisms （SNPs） in Hordoindoline a （hina） gene and its relationship with hardness index among North American harley cultivars were investigated. The hina genes in 36 cultivars were sequenced for the SNP analyses, and 17 sets of SNP primers were designed to detect the SNP variations of hina locus in 92 North American barley cultivars. SNP detection indicated that there were four haplotypes in the hina genes of 92 barley cultivars, and haplotype 01 and 02 were shared by 68 and 14 cultivars, respectively, suggesting that there was a very limited diversity in hina genes among North American barley cultivars. Despite the wide range in hardness exists in 92 barley cultivars, however, unlike wheat, where a clear relationship has been demonstrated between a number of SNPs in the wheat hardness genes and quality （soft or hard wheat）, there was no such relationship for barley. The genotypes used in this study demonstrated that there was a low level of polymorphism in hina gene in North American barley cultivars and these polymorphisms had no impact on grain hardness. 相似文献
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The hina gene encodes a HINA protein in seeds of barley （Hordeum vulgare）, which was known to affect the grain hardness. 171 hina gene sequences from Tibetan wild barley accessions and worldwide were characterized. Across 1 452 nucleotides of 171 hina genes, 152 SNPs were detected, giving an average frequency of one SNP per 9.5 bases. There were 93 singleton variable sites （the nucleotide polymorphism only observed in a single accession）, 59 polymorphic sites （the polymorphisms found in two or more accessions） and 8 indels. A total of 18 haplotypes were defined, and most of the barley accessions shared one gene haplotype. H. spontaneum had a wider haplotype distribution. Through the analysis of median-joining network of the 18 haplotypes, 4 haplotype groups were found, which were testified by neighbor-joining tree based on the complete sequence alignment. Extremely low level of hina gene diversity was observed in Tibetan wild barley accessions, indicating that Tibet is unlikely a center of origin for cultivated barley. 相似文献
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The plant hormone abscisic acid (ABA) regulates many important physiological and developmental processes in plants.The objective of this study was to clone the ABA 8'-hydroxylase gene in common wheat. In the present study, we used the cDNA sequence of barley HvCYP707A1 gene (GenBank accession no. AB239299) as a probe for BLAST search against the common wheat (Triticum aestivum L.) EST database in GenBank. All wheat ESTs sharing high similarity with the reference gene were subjected to contig assembly. Primers were designed based on the constructed contigs to clone the wheat CYP707A1 gene, designated as TaCYP707AI. The genomic DNA sequence of TaCYP707A1 gene comprised five exons and four introns, with a size of 2 225 bp. The corresponding cDNA sequence of TaCYP707A1 was 1 737 bp,containing an open reading frame (ORF) of 1 431 bp, a 42-bp 5'-untranslated region (UTR) and a 264-bp 3'UTR, with 94.9% of identical sequences to HvCYP707A1 gene (AB239299). The neighbor joining tree indicated that the deduced amino acid sequences of TaCYP707A1 gene was highly similar to those of barley and rice. The TaCYP707A1 gene was located on chromosome 6BL using a set of Chinese Spring nullisomic-tetrasomic lines and ditelosomic line 6BS. These results will be of high importance in understanding of molecular mechanism of ABA catabolism. 相似文献
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为初步探究暗纹东方鲀sox9基因在性腺发育和性别分化过程中的作用,通过设计兼并引物、RACE及荧光定量PCR技术,成功克隆出暗纹东方鲀sox9基因的cDNA全长序列,并分析其相应的生物信息学特征及其在雌雄个体的组织表达水平。结果表明:sox9a基因cDNA全长为1 248 bp(NCBI登录号:MH218818),包括684 bp的ORF,编码227个氨基酸,297 bp的5′UTR和267 bp的3′UTR。sox9b基因全长为1 941 bp(NCBI登录号:MH218819),包括1 470 bp的ORF,编码489个氨基酸,306 bp的5′UTR和165 bp的3′UTR。同源性和系统发育分析结果表明暗纹东方鲀sox9基因与红鳍东方鲀同源性最高,亲缘关系最近。氨基酸多重序列比对结果显示两个Sox9氨基酸序列的HMG box结构域在哺乳动物和鱼类中均高度保守。荧光定量PCR分析显示两个sox9基因普遍存在于雌鱼和雄鱼的各个组织中,且均在雌鱼下丘脑中表达量最高,在精巢中有少量表达,卵巢中几乎不表达。总体来说,除少数组织外,两个sox9基因在雌鱼组织中的表达量普遍高于雄鱼。本研究为了解暗纹东方鲀sox9的遗传特性及相关的生理功能,探索性别分化与性腺发育的分子调控机制提供理论依据。 相似文献
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以普通小麦品种济麦21为材料,根据GenBank中已发表的LOX1基因序列设计引物,利用RTPCR技术获得一个小麦LOX1的基因片段,并对该片段进行测序和生物信息学分析。结果表明:克隆到的基因片段为384bp(GenBank登录号JX126806),GC含量为64.84%,推导氨基酸残基为127个,分子质量为11.15ku,等电点(pI)为4.93,与杜伦小麦、大麦、高粱、水稻的同源性分别为99.7%、94.8%、73.7%、56.2%。进化树分析表明,六倍体普通小麦LOX1基因与杜伦小麦LOX1基因(GenBank登录号HM126468)亲缘关系较近,先聚为一支,然后再与大麦的LOX1基因(GenBank登录号L35931.1)、高粱的LOX1基因(GenBank登录号GQ369443)聚为一类,最终与亲缘关系较远的水稻LOX1基因(GenBank登录号EU267789)聚类,这与物种亲缘关系的远近基本一致,表明所得LOX1基因片段可作为研究物种亲缘关系或遗传距离的参考标记之一。 相似文献
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蔗糖:蔗糖1-果糖基转移酶基因1-SST在植物逆境胁迫反应中起重要作用。为了挖掘华山新麦草(Psathyrostachys huashanica Keng)抗非生物胁迫关键功能基因,以华山新麦草叶片为材料,利用RT-PCR结合RACE技术克隆1-SST基因的全长cDNA,命名为Ph-1-SST,登录号为KX761897。通过PCR法扩增其gDNA序列,测序结果表明该基因的gDNA和cDNA序列长度分别为3 344bp和2 001bp,编码666个氨基酸残基,序列结构分析结果表明该基因含4个外显子3个内含子,预测该蛋白相对分子质量为72.9ku,理论等电点为4.87。序列比对显示,该基因与大麦1-SST基因编码蛋白的相似性最高(90%),为糖基水解酶32家族成员。对1-SST氨基酸序列的系统进化树分析显示,Ph-1-SST及其同源蛋白位于不同分支,初步推断此全长cDNA是华山新麦草中编码蔗糖:蔗糖1-果糖基转移酶的一个新基因。结果为作物非生物胁迫改良提供重要基因资源。 相似文献
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根据小麦TaSIM基因的cDNA序列设计引物,采用PCR技术从小麦中克隆TaSIM基因的DNA序列,采用半定量RT-PCR方法研究TaSIM基因在不同组织中的表达。结果表明:TaSIM编码区DNA序列长度为2 355bp,包含2个外显子和1个内含子。分析发现内含子富含AT,在内含子中发现2个MYB转录因子结合位点。半定量RT-PCR检测表明,TaSIM基因在不同组织中均有表达,在雄蕊中的表达量最高。TaSIM表达量依次为雄蕊>雌蕊>根>茎>叶。 相似文献
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为获得马鹿(Cervus elaphus)Ghrelin全长cDNA序列并进行序列比较和分析,本试验以马鹿皱胃黏膜组织内提取的总RNA为模板,通过RT-PCR、RACE和基因克隆等技术进行克隆。结果表明:获得长度为539bp的马鹿cDNA全序列(GenBank accession no.KX857494),其中包括46bp的5′非编码区(5′UTR),351bp的开放阅读框(ORF),编码116个氨基酸残基的前原Ghrelin(preproghrelin),128bp的3′非编码区(3′UTR)和poly(A)14;Ghrelin的氨基酸同源性分析表明,马鹿Ghrelin cDNA全序列与驯鹿、梅花鹿、山羊、绵羊、牛等物种间的同源性较高,进化树分析与其亲缘关系远近一致。 相似文献
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为获得山葡萄LDOX基因的全长序列,采用RT-PCR与SMART RACE技术克隆LDOX基因,并对该基因进行生物信息学分析。结果显示,山葡萄LDOX基因全长1 353bp,其中开放阅读框(ORF)为1 068bp,编码355个氨基酸,氨基酸序列的分子质量为40.19ku,等电点为5.61;VAmLDOX基因(GenBank登陆号:FJ645769)属于双加氧酶基因家族,不含信号肽,VAmLDOX蛋白属于不稳定亲水蛋白,二级结构中随机卷曲含量最高;山葡萄VAmLDOX氨基酸序列与欧亚种葡萄、苹果、大豆、三花龙胆和紫苏等的同源性系数分别为99%、81%、80%、77%和75%;半定量RT-PCR分析显示,在山葡萄果实着色过程中,VAmLDOX在不同时期的果皮中均有表达,在转色期的叶片、茎、果肉中也均有表达,且表达量相近。 相似文献
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为研究柔嫩艾美耳球虫不同地理株的线粒体细胞色素c氧化酶第1亚基(cox1)基因与球虫种群之间的遗传关系,以安徽3个地区的3株柔嫩艾美耳球虫为研究对象,通过卵囊的分离与纯化获得纯种虫株,然后应用PCR技术对柔嫩艾美球虫3个地理株的cox1序列进行扩增,并与GenBank上登录的鸡柔嫩艾美耳球虫、巨型艾美耳球虫、布氏艾美耳球虫和堆形艾美耳球虫虫株的相应序列进行比对分析。结果显示,每个虫株都成功扩增出800 bp左右的cox1部分有效序列(pcox1),与柔嫩艾美耳球虫pcox1序列种内无差异,pcox1相应序列种间的差异程度为9.9%～14.9%。表明柔嫩艾美耳球虫的cox1序列可作为艾美耳球虫种间遗传变异研究的标记,为进一步研究艾美耳球虫的群体遗传学特性和耐药性奠定了基础。 相似文献
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为获得梅花鹿Ghrelin cDNA全序列,以梅花鹿皱胃黏膜上皮组织提取的总RNA为模板,通过RT-PCR和RACE法克隆了梅花鹿皱胃中Ghrelin基因cDNA的全序列。结果表明梅花鹿Ghrelin cDNA序列全长为539bp,其中5’非翻译区(5’UTR)为46bp,3’UTR为128bp,开放阅读框(ORF)为351bp,该ORF编码116个氨基酸残基。将梅花鹿Ghrelin基因的cDNA与人和其他动物的Ghrelin相比,发现:梅花鹿Ghrelin与驯鹿、山羊、绵羊和牛的同源性达90.4%~99.1%;与恒河猴、人、猪、犬的同源性达76.6%~66.9%;与鸡和野鸽的同源性分别为36.4%和35.4%。研究表明Ghrelin的结构具有明显的种属特异性,因此Ghrelin在反刍动物体内可能有着重要的生理功能。 相似文献