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小麦AGPase胞质型基因LSUI的克隆及正义、反义与RNAi干扰载体的构建 被引量:4
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作者 康国章 张孟琴 +2 位作者 官春云 郭天财 朱云集 《浙江大学学报(农业与生命科学版)》 CAS CSCD 北大核心 2007年第6期607-615,共9页
采用RT-PCR方法从小麦品种豫教2号的发育籽粒中克隆出AGPase胞质型大亚基基因(large subunit,LSU I)的cDNA全长(1947 bp,GenBank No.DQ839506),同源性比较结果表明,与GenBank上已报道的LSU I基因同源性达99%,虽与其有2处碱基不同(与Z21... 采用RT-PCR方法从小麦品种豫教2号的发育籽粒中克隆出AGPase胞质型大亚基基因(large subunit,LSU I)的cDNA全长(1947 bp,GenBank No.DQ839506),同源性比较结果表明,与GenBank上已报道的LSU I基因同源性达99%,虽与其有2处碱基不同(与Z21969相比,分别在851 bp处的T变为C和926 bp处的G变为A),但他们的氨基酸序列相同,故不影响其功能.以pBI121质粒为基础,通过中间载体pBSK,构建了由35S启动子调控的LSU I基因的正义表达载体pBI121LSUⅠS;将该基因反向置于pBI121质粒的CaMV35S启动子之后,构建了LSU I的反义表达载体pBI121LSUⅠA.同时还克隆出LSU I基因的250 bp片段,以pFGC5941质粒为基础,构建了RNAi干扰载体pFGCLsuⅠ,这为研究该基因的功能打下了很好的基础. 展开更多
关键词 普通小麦 AGPase胞质型大亚基 表达载体
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实验感染大、小鼠和临床疑似病例肺孢子虫mt LSU rRNA基因检测 被引量:2
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作者 冯宪敏 李自慧 +2 位作者 卢思奇 张帆 王凤云 《寄生虫与医学昆虫学报》 CAS 2006年第3期129-134,共6页
为探讨PCR技术对实验感染大、小鼠肺孢子虫肺炎(PCP)和临床疑似病例诊断的可行性。用皮下注射地塞米松法诱导SD大鼠和ICR小鼠PCP并收集临床疑似病例24h深部痰液。受试动物解剖后,制备肺印片,经瑞姬氏复合染色镜检确定肺孢子虫(Pc)感染... 为探讨PCR技术对实验感染大、小鼠肺孢子虫肺炎(PCP)和临床疑似病例诊断的可行性。用皮下注射地塞米松法诱导SD大鼠和ICR小鼠PCP并收集临床疑似病例24h深部痰液。受试动物解剖后,制备肺印片,经瑞姬氏复合染色镜检确定肺孢子虫(Pc)感染的阳性率;同时,以针对虫体mtLSUrRNA基因设计的引物,PCR扩增鼠肺组织和支气管灌洗液(BALF),以及痰液样本中的DNA;比较实验动物样本镜检和PCR两种方法的检测结果;测定PCR的敏感性和特异性。结果表明,SD大鼠和ICR小鼠肺印片的阳性率分别为68.8%(1116)和85.7%(1821),肺组织PCR检测的阳性率分别为75%(1216)和80.9%(1721),肺泡灌洗液PCR的阳性率分别为81.2%(1316)和23.8%(521)。肺印片镜检和PCR技术检测结果无显著性差别;可见,PCR对虫体mtLSUrRNA基因的检测具有显著的特异性和敏感性,至少可以检测出0.6pg的肺孢子虫DNA;6例临床疑似病例样本中2例显示肺孢子虫PCR阳性反应。 展开更多
关键词 肺孢子虫 肺孢子虫肺炎 MT lsu RRNA基因 临床疑似病例 基因检测
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Molecular Phylogenetic Lineage of Plagiopogon and Askenasia(Protozoa, Ciliophora) Revealed by Their Gene Sequences
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作者 LIU An YI Zhenzhen +3 位作者 LIN Xiaofeng HU Xiaozhong Saleh A.AL-FARRAJ Khaled A.S.AL-RASHEID 《Journal of Ocean University of China》 SCIE CAS 2015年第4期724-730,共7页
Prostomates and haptorians are two basal groups of ciliates with limited morphological characteristics available for taxonomy. Morphologically, the structures used to identify prostomates and haptorians are similar or... Prostomates and haptorians are two basal groups of ciliates with limited morphological characteristics available for taxonomy. Morphologically, the structures used to identify prostomates and haptorians are similar or even identical, which generate heavy taxonomic and phylogenetic confusion. In present work, phylogenetic positions lineage of two rare genera, Plagiopogon and Askenasia, were investigated. Three genes including small subunit ribosomal RNA gene(hereafter SSU r DNA), internal transcribed spacer region(ITS region), and large subunit ribosomal RNA gene(LSU r DNA) were analyzed, 10 new sequences five species each. Our findings included 1) class Prostomatea and order Haptorida are multiphyletic; 2) it may not be appropriate to place order Cyclotrichiida in subclass Haptoria, and the systematic lineage of order Cyclotrichiida needs to be verified further; 3) genus Plagiopogon branches consistently within a clade covering most prostomes and is basal of clade Colepidae, implying its close lineage to Prostomatea; and 4) Askenasia is phylogenetically distant from the subclass Haptoria but close to classes Prostomatea, Plagiopylea and Oligohymenophorea. We supposed that the toxicyst of Askenasia may be close to taxa of prostomes instead of haptorians, and the dorsal brush is a more typical morphological characteristics of haptorians than toxicysts. 展开更多
关键词 分子系统发育 基因序列 纤毛虫 原生动物 核糖体RNA基因 谱系 显示 形态特征
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高产紫杉醇内生真菌J11的鉴定 被引量:7
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作者 刘明志 段中岗 +1 位作者 吕镇城 刘春霞 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2014年第10期1031-1038,共8页
从南方红豆杉(Taxus chinensis var.mairei)的幼茎中分离出一株产紫杉醇内生真菌J11.菌株J11的发酵提取物经高分辨质谱分析,证实J11菌株可产紫杉醇.提取该菌株的基因组DNA,扩增核糖体internal transcribed spacer(ITS)和28S核糖体large ... 从南方红豆杉(Taxus chinensis var.mairei)的幼茎中分离出一株产紫杉醇内生真菌J11.菌株J11的发酵提取物经高分辨质谱分析,证实J11菌株可产紫杉醇.提取该菌株的基因组DNA,扩增核糖体internal transcribed spacer(ITS)和28S核糖体large subunit rRNA gene(LSU)序列,经测序获得该菌的ITS序列和LSU序列.序列比对和检索结果表明,J11菌株为葡萄座腔菌(Botryosphaeria ssp.)属中的一个新菌株.形态学鉴定符合葡萄座腔菌属特征,高效液相色谱分析表明,J11菌株的紫杉醇含量约为615.1μg/L.本研究首次证实葡萄座腔菌J11是一株高产紫杉醇野生型菌株,具有潜在的应用前景. 展开更多
关键词 内生真菌 紫杉醇 葡萄座腔菌 核糖体内部转录间隔序列 28S核糖体大亚基基因
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福建古田红曲生产用红曲霉菌主要种类的鉴别 被引量:10
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作者 李志强 刘颖 +1 位作者 林风 吴丽云 《食品与发酵工业》 CAS CSCD 北大核心 2017年第5期64-69,共6页
以购自中国工业微生物菌种保藏管理中心(CICC)的5株红曲霉菌为对照,对福建省微生物研究所红曲霉菌资源库中收藏自福建省古田县的60株红曲霉菌(Monascus sp.)进行分类鉴定研究。首先根据不同菌株在麦芽汁琼脂(Wa)、察氏酵母提取物琼脂(C... 以购自中国工业微生物菌种保藏管理中心(CICC)的5株红曲霉菌为对照,对福建省微生物研究所红曲霉菌资源库中收藏自福建省古田县的60株红曲霉菌(Monascus sp.)进行分类鉴定研究。首先根据不同菌株在麦芽汁琼脂(Wa)、察氏酵母提取物琼脂(CYA)和甘油硝酸盐琼脂(G25N)培养基上的菌落形态特征进行初步分类,进而基于核糖体DNA内转录间隔区(rDNA internal transcribed spacer,rDNA ITS)、核糖体大亚基(28S nuclear ribosomal large subunit rRNA gene,LSU)和核糖体小亚基(18S nuclear ribosomal small subunit r RNA gene,SSU)的基因片段的测序结果,在Gen Bank中通过BLAST进行分子生物学方面的鉴定研究,并用ITS序列构建系统发育树。通过形态特征及3个基因序列的比对,认为所选取的60株古田红曲霉菌分别归属红色红曲霉菌(M.ruber)、丛毛红曲霉菌(M.pilosus)或紫色红曲霉菌(M.purpureus)3个种。 展开更多
关键词 红曲霉菌 ITS lsu SSU 鉴定
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真菌共祖起源的分子证据 被引量:4
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作者 付少彬 魏江春 《菌物学报》 CAS CSCD 北大核心 2008年第2期217-223,共7页
以真菌界、动物界和植物界中的19种各类生物作为代表,从具有进化速率不同区段结构的核糖体核酸基因大亚基全序列的C3、C9及C11三个高度保守区段中鉴定出了4条真菌所共有的核苷酸序列,亦即真菌界所特有的核苷酸序列。从其保守区段C5中鉴... 以真菌界、动物界和植物界中的19种各类生物作为代表,从具有进化速率不同区段结构的核糖体核酸基因大亚基全序列的C3、C9及C11三个高度保守区段中鉴定出了4条真菌所共有的核苷酸序列,亦即真菌界所特有的核苷酸序列。从其保守区段C5中鉴定出了1条子囊菌门所特有的保守序列;从保守区段C7中鉴定出了1条担子菌门所特有的保守序列。研究结果为各类真菌起源于一个共同祖先提供了分子证据。用于研究的美味石耳的核糖体核酸基因大亚基全序列是由本实验室所提供;其他18种核糖体核酸基因大亚基全序列来自GenBank。 展开更多
关键词 美味石耳 子囊菌门 担子菌门 核糖体核酸基因 特有序列
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厌氧真菌系统发育分类及多样性研究进展 被引量:1
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作者 张亚伟 王月红 +1 位作者 张元庆 刘强 《动物营养学报》 CAS CSCD 北大核心 2022年第2期700-709,共10页
厌氧真菌[新毛孢菌门(Neocallimastigomycota)]是哺乳草食动物消化道内常见的微生物之一,在木质纤维性饲料原料的降解过程中起着至关重要的作用。了解厌氧真菌个体间的系统发育关系对于进一步了解它们的进化史,以及驱动它们在不同类型... 厌氧真菌[新毛孢菌门(Neocallimastigomycota)]是哺乳草食动物消化道内常见的微生物之一,在木质纤维性饲料原料的降解过程中起着至关重要的作用。了解厌氧真菌个体间的系统发育关系对于进一步了解它们的进化史,以及驱动它们在不同类型宿主内部及之间的生态位专门化的因素等方面至关重要。本文从系统发育分类学发展史、低等级的分类学多样性及相关分子层面的研究方法对近年来有关厌氧真菌系统发育分类及多样性的研究进行了概述,以期对相关的研究工作提供一定的参考和帮助。 展开更多
关键词 草食动物 厌氧真菌 系统发育 内部转录间隔区(ITS) 大亚基rRNA基因(lsu)
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DNA barcode assessment of Ceramiales(Rhodophyta) in the intertidal zone of the northwestern Yellow Sea 被引量:1
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作者 杜国英 吴菲菲 +2 位作者 郭皓 薛红凡 茅云翔 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2015年第3期685-695,共11页
A total of 142 specimens of Ceramiales(Rhodophyta) were collected each month from October 2011 to November 2012 in the intertidal zone of the northwestern Yellow Sea. These specimens covered 21 species,14 genera,and f... A total of 142 specimens of Ceramiales(Rhodophyta) were collected each month from October 2011 to November 2012 in the intertidal zone of the northwestern Yellow Sea. These specimens covered 21 species,14 genera,and four families. Cluster analyses show that the specimens had a high diversity for the three DNA markers,namely,partial large subunit r RNA gene(LSU),universal plastid amplicon(UPA),and partial mitochondrial cytochrome c oxidase subunit I gene(COI). No intraspecific divergence was found in our collection for these markers,except for a 1–3 bp divergence in the COI of Ceramium kondoi,Symphyocladia latiuscula,and Neosiphonia japonica. Because short DNA markers were used,the phylogenetic relationships of higher taxonomic levels were hard to evaluate with poor branch support. More than half species of our collection failed to find their matched sequences owing to shortage information of DNA barcodes for macroalgae in Gen Bank or BOLD(Barcode of Life Data) Systems. Three specimens were presumed as H eterosiphonia crispella by cluster analyses on DNA barcodes assisted by morphological identification,which was the first record in the investigated area,implying that it might be a cryptic or invasive species in the coastal area of northwestern Yellow Sea. In the neighbor-joining trees of all three DNA markers,H eterosiphonia japonica converged with D asya spp. and was distant from the other Heterosiphonia spp.,implying that H. japonica had affinities to the genus Dasya. The LSU and UPA markers amplified and sequenced easier than the COI marker across the Ceramiales species,but the COI had a higher ability to discriminate between species. 展开更多
关键词 DNA标记 条形码 北黄海 潮间带 红藻门 细胞色素C氧化酶 GENBANK 评估
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mtLSU-巢式PCR检测实验大鼠卡氏肺孢子虫的实验研究
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作者 张小娟 杨芳芳 +3 位作者 欧阳颐 王鸽 黎学铭 林睿 《热带医学杂志》 CAS 2011年第4期372-374,432,共4页
目的对mtLSU-巢式PCR方法检测大鼠卡氏肺孢子虫的应用价值以及基因序列进行评价。方法采用地塞米松免疫抑制法诱导大鼠感染肺孢子虫;实验组10只,对照组1只;诱导至第7周时收集实验组及对照组大鼠肺组织和支气管肺泡灌洗液(BALF)标本,采用... 目的对mtLSU-巢式PCR方法检测大鼠卡氏肺孢子虫的应用价值以及基因序列进行评价。方法采用地塞米松免疫抑制法诱导大鼠感染肺孢子虫;实验组10只,对照组1只;诱导至第7周时收集实验组及对照组大鼠肺组织和支气管肺泡灌洗液(BALF)标本,采用mtLSU-巢式PCR方法对人源与鼠源肺孢子虫共有的基因进行扩增和序列测定,同时采用镜检法对实验组大鼠肺组织和肺泡灌洗液标本进行检测,评估两种方法的敏感性。结果采用mtLSU-巢式PCR方法对实验感染大鼠肺组织和BAL进行检测,卡氏肺孢子虫DNA阳性率分别为100%(10/10)、90%(9/10)。而GMS染色镜检法检测的阳性率分别为80%(8/10)、60%(6/10)。所测Wistar大鼠卡氏肺孢子虫mtLSU基因序列长度为155bp,与GenBank的大鼠源肺孢子虫(U20170)及人源肺孢子虫(DQ473446)同源性均为100%(154/154、155/155)。结论 mtLSU-巢式PCR方法应用于大鼠卡氏肺孢子虫检测敏感性高,特异性强;获得与人源耶氏肺孢子虫相同的Wistar大鼠卡氏肺孢子虫mtLSU的基因序列。 展开更多
关键词 卡氏肺孢子虫 线粒体核糖体大亚基RNA基因(mtlsu) 巢式PCR 基因序列
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Taxonomy and phylogeny of the section Chaetoceros(Chaetocerotaceae,Bacillariophyta),with description of two new species
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作者 Xudan LU Mengyi ZHAI +1 位作者 Nina LUNDHOLM Yang LI 《Journal of Oceanology and Limnology》 SCIE CAS 2024年第4期1286-1311,共26页
Chaetoceros Ehrenberg is one of the most diverse genera of planktonic diatoms.The species in section Chaetoceros are characterized by cells and setae having numerous chloroplasts and being widely distributed.However,t... Chaetoceros Ehrenberg is one of the most diverse genera of planktonic diatoms.The species in section Chaetoceros are characterized by cells and setae having numerous chloroplasts and being widely distributed.However,the delimitations of some species are problematic because of limited morphological information in the classical descriptions.Monoclonal strains of the section Chaetoceros were established,morphological features were studied using light and electron microscopy,and the hypervariable D 1-D 3 region of the nuclear ribosomal large subunit gene was sequenced to address phylogenetic relationships.Fifteen species belonging to the section Chaetoceros were recorded,including two new species,C.hainanensis sp.nov.and C.tridiscus sp.nov.Chaetoceros hainanensis was characterized by straight chains,narrowly lanceolate to hexagonal apertures,sibling setae diverging in nearly right angles,stipule-shaped spines on terminal setae and arrowhead-shaped spines on intercalary setae.C.tridiscus had short straight chains,narrowly lanceolate apertures,arrowhead-shaped spines and circular poroids arranged in a grid pattern on terminal and intercalary setae.The phylogenetic analyses revealed six groups formed by 19 species within the section Chaetoceros,which was found to be monophyletic.The subdivision of the section is still not well understood.The morphological characters within each group varied considerably and molecular information on more species are needed to enrich the phylogenetic profiling. 展开更多
关键词 morphology large subunit ribosomal RNA encoding gene(lsu) section Chaetoceros Chaetoceros hainanensis Chaetoceros tridiscus
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