期刊文献+
共找到27篇文章
< 1 2 >
每页显示 20 50 100
中国棉铃虫核多角体病毒VHA273毒株DNA的特性 被引量:6
1
作者 吴柏春 王行国 《病毒学报》 CAS CSCD 北大核心 1990年第4期352-357,共6页
VHA273毒株纯多角体经碱解、酶解及饱和酚一步法和二步法抽提的基因组,具有典型的DNA吸收光谱。吲哚法亦确证为DNA,无RNA。超离心沉降分析,扫描出主峰沉降常数为26S,次峰9.7S。热变性温度法测得Tm=89.6℃,计算(G+C)%=49.53。电镜观察DN... VHA273毒株纯多角体经碱解、酶解及饱和酚一步法和二步法抽提的基因组,具有典型的DNA吸收光谱。吲哚法亦确证为DNA,无RNA。超离心沉降分析,扫描出主峰沉降常数为26S,次峰9.7S。热变性温度法测得Tm=89.6℃,计算(G+C)%=49.53。电镜观察DNA呈3种典型形态。大的闭环DNA平均长度29.8μ,计算得分子量是61.7×10~6;有少量极小闭环DNA存在。0.7%琼脂糖凝胶电泳显示,DNA主带清晰浓重,发现还有5条微弱次带,约2.5~3.7kb。该DNA用4种限制性核酸内切酶Bg1 Ⅱ、BamHⅠ、EcoRⅠ和HindⅢ分别酶切成11、8、14和13条带。用片段积加法求出的分子量彼此很接近,平均为67.0×10~6道尔顿。酶切图谱显示,5条弱带对4种内切酶概不敏感。 展开更多
关键词 中国棉铃虫 核多角体病毒 dna特性
下载PDF
凝胶迁移阻滞法分析人类端粒酶的DNA结合特性 被引量:1
2
作者 周俊宜 周俊梅 +5 位作者 谢金卫 周世豪 杜贻鹏 谢奇峰 陶莎 罗超权 《中国老年学杂志》 CAS CSCD 北大核心 2005年第5期582-584,共3页
目的采用凝胶迁移阻滞法分析研究人类端粒酶DNA结合特性。方法以地高辛标记的人端粒序列寡核苷酸与人端粒酶提取物进行结合反应,以未参与结合反应的单纯标记寡核苷酸作为参照。实验所设的3组样品中,第一组和第二组标记寡核苷酸加端粒酶... 目的采用凝胶迁移阻滞法分析研究人类端粒酶DNA结合特性。方法以地高辛标记的人端粒序列寡核苷酸与人端粒酶提取物进行结合反应,以未参与结合反应的单纯标记寡核苷酸作为参照。实验所设的3组样品中,第一组和第二组标记寡核苷酸加端粒酶,第一组显示一条阻滞性条带,第二组显示两条条带,前方的一条与第一组条带平行,为单纯探针条带,后方的一条迁移率被阻滞,应为探针与纯化的蛋白复合体相结合的结果;第三组为标记寡核苷酸加端粒酶阴性洗涤液,只显示单一探针条带。结果只有人端粒序列寡核苷酸与人端粒酶提取物结合反应管显示特异的凝胶迁移阻滞性电泳条带。结论人端粒酶具有和端粒DNA片段结合的特性。 展开更多
关键词 端粒酶 凝胶迁移阻滞法 dna结合特性
下载PDF
热变性法测定红色毛癣菌核中DNA鸟嘌呤加胞嘧啶含量的研究
3
作者 滕传远 吴绍熙 《中国医学科学院学报》 CAS CSCD 北大核心 1989年第4期313-316,共4页
DNA中鸟嘌呤加胞嘧啶克分子百分比含量(G+Cmol%)值是一种间接反映两个物种核中DNA序列相似性的遗传指标之一.自从Lee和Belozersky把G+Cmol%值用作细菌分类鉴定遗传指标以后,G+Cmol%值在微生物分类鉴定方面已得到广泛应用。在真菌方面... DNA中鸟嘌呤加胞嘧啶克分子百分比含量(G+Cmol%)值是一种间接反映两个物种核中DNA序列相似性的遗传指标之一.自从Lee和Belozersky把G+Cmol%值用作细菌分类鉴定遗传指标以后,G+Cmol%值在微生物分类鉴定方面已得到广泛应用。在真菌方面,国外于60年代已有报道。 展开更多
关键词 热变性法 红公抟癣菌 dna特性
下载PDF
Changes in Physiological Properties and Respiratory Pathway of the New Lines of Wheat Introduced Exogenous DNA Under Salt Stress 被引量:6
4
作者 孔英珍 周功克 +1 位作者 崔凯荣 王亚馥 《Acta Botanica Sinica》 CSCD 2001年第3期249-255,共7页
New lines of wheat ( Triticum aestivum L.) was obtained by introducing the DNA of sorghum (Sorghum vulgare Pers.) into wheat cultivar 'Longchun 13'. The changes of respiratory pathway, contents of protein, Na+... New lines of wheat ( Triticum aestivum L.) was obtained by introducing the DNA of sorghum (Sorghum vulgare Pers.) into wheat cultivar 'Longchun 13'. The changes of respiratory pathway, contents of protein, Na+ and K+ in the leaves and roots of the new lines of wheat under salt stress were determined and compared with the control cultivar, 'Longchun 13'. The decrease of the content of K+ was observed with the increase of NaCl concentrations, but the decrease was more in the control than that in the new lines, and more in roots than in leaves. Content of proline and Na+ in both two wheats lines increased greatly, but the former increased more significantly in the new lines and the latter more significantly in control both in leaves and roots. The operation of the cyanide-resistant pathway of respiration was enhanced at different degrees after salt stress and it increased much more in roots and leaves of the control plant than that in the new lines, but the cytochrome pathway of electron transport was still the main one consistently. The possible significance of these changes was discussed. 展开更多
关键词 salt stress salt-tolerant wheat respiratory pathway
下载PDF
Characterization of Two Groups of Low_copy and Specific DNA Sequences Isolated from Chromosome 7B of Common Wheat 被引量:2
5
作者 刘振兰 董玉柱 刘宝 《Acta Botanica Sinica》 CSCD 2002年第8期946-950,共5页
Recent work revealed that, in the genomes of polyploid wheat, there exists a class of low_copy and chromosome_specific sequences that are labile upon polyploid formation. This class of sequences was proposed to play ... Recent work revealed that, in the genomes of polyploid wheat, there exists a class of low_copy and chromosome_specific sequences that are labile upon polyploid formation. This class of sequences was proposed to play a critical role in the stabilization and establishment of nascent plant polyploids as new species. To further study this issue, five wheat chromosome 7B_specific sequences, isolated from common wheat (Triticum aestivum L.) by chromosome microdissection, were characterized. The sequences were studied by genomic Southern hybridizations on a collection of polyploid wheats and their diploid progenitors. Four sequences hybridized to all polyploid species, but at the diploid level to only species closely related to the B_genome of polyploid wheat. This indicates that these sequences originated with the divergence of the diploid species, and was then vertically transmitted to polyploids. One sequence hybridized to all species at both the diploid and polyploid levels, suggesting its elimination after the polyploid wheat formation. The hybridization of this sequence to two synthetic polyploid wheats indicated that sequence elimination is a rapid event and probably related to methylation status of the sequence. Based on the above results, we suggest that selective changes of low_copy sequences occur rapidly after polyploid formation, which may contribute to the differentiation of chromosomes in newly formed allopolyploid wheats. 展开更多
关键词 polyploid wheat chromosome_specific dna sequences sequence elimination dna methylation genome evolution
下载PDF
Studies of Site Specific DNA Binding of Small Peptides by Competitive Assays with Hoechst 33258
6
作者 杨铭 朱树梅 +2 位作者 黄艳萍 胡齐悦 王夔 《Journal of Chinese Pharmaceutical Sciences》 CAS 1996年第3期141-146,共6页
With a view to finding out precisely how small peptides recognize a particular binding site of DNA, we have accomplished DNA binding studies of two peptides, H-Tyr-Arg-OH (YR) and H-Gly-Gly-His-OH (GGH) by using measu... With a view to finding out precisely how small peptides recognize a particular binding site of DNA, we have accomplished DNA binding studies of two peptides, H-Tyr-Arg-OH (YR) and H-Gly-Gly-His-OH (GGH) by using measurements in comparison with the binding between DNA and Hoechst 33258. The inhibition mode by YR and GGH to DNA binding of Hoechst 33258 was analyzed by Lineweaver-Burk plot which shows the plot of typical competitive inhibition at concentration of Hoechst 33258 from 3.66 ( 10-9 mol / L to 1.09 ( 10-8 mol / L. And it is concluded that YR binds to DNA in its minor groove (AT rich regions) with a binding constant K = 1.02 ( 108 (mol / L)-1. The GGH(s specificity is reduced at high concentration because it can also bind GC base pair. 展开更多
关键词 dna PEPTIDE Competitive inhibitor Site specific binding
下载PDF
Fluorescent vital staining of plant sexual cell nuclei with DNA-specific fluorochromes and its application in gametoplast fusion 被引量:2
7
作者 YANGHONGYUAN XINLIWU 《Cell Research》 SCIE CAS CSCD 1993年第2期121-130,共10页
DNA-binding fluorochromes are often used for vital staining of plant cell nuclei. However, it is not always sure whether the cells after staining still remain in living state. We chose several criteria to estimate the... DNA-binding fluorochromes are often used for vital staining of plant cell nuclei. However, it is not always sure whether the cells after staining still remain in living state. We chose several criteria to estimate the validity of real vital staining for sexual cell nuclei. These were: the cytoplasmic streaming in pollen tubes whose nuclei were stained, the simultaneous visualization of fluo-rochromatic reaction and nucleus staining in isolated generative cells, and the capability of isolated, prestained generative or sperm cells to fuse with other protoplasts. The results confirmed that 4',6-diamidino-2-phenylindole (DAPI), Hoechst 33258 and mithramycin could be used as real vital stains, though their efficiency varied from case to case; among them DAPI showed best effect. The fluorescent vital staining technique offered a useful means fori-dentification and selection of heterokaryons in gametoplast manipulation studies. 展开更多
关键词 fluorescent vital staining dna-specific fluorochrome generative cell sperm cell gametoplast fusion.
下载PDF
Statistical properties of nucleotide clusters in DNA sequences
8
作者 成军 章林溪 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE EI CAS CSCD 2005年第5期408-412,共5页
Using the complete genome of Plasmodium falciparum 3D7 which has 14 chromosomes as an example, we have examined the distribution functions for the amount of C or G and A or T consecutively and non-overlapping blocks o... Using the complete genome of Plasmodium falciparum 3D7 which has 14 chromosomes as an example, we have examined the distribution functions for the amount of C or G and A or T consecutively and non-overlapping blocks of m bases in this system. The function P(S) about the number of the consecutive C-G or A-T content cluster conforms to the relation P(S)∝e? ; αs values of the scaling exponent αCG are much larger than αAT; and αAT of 14 chromosomes are hardly changed, whereas αCG of 14 chromosomes have a number of fluctuations. We found maximum value of A-T cluster size is much larger than C-G, which implies the existence of large A-T cluster. Our study of the width function ξ(m) of cluster C-G content showed that follows good power law ξ(m)∝m?γ. The average γ for 14 chromosomes is 0.931. These investigations provide some insight into the nucleotide clusters of DNA sequences, and help us understand other properties of DNA sequences. 展开更多
关键词 dna sequence Plasmodium falciparum 3D7 Nucleotide clusters Power law
下载PDF
Ashkin-Teller Formalism for Elastic Response of DNA Molecule to External Force and Torque
9
作者 CHANG Zhe WANG Ping ZHENG Ying-Hong 《Communications in Theoretical Physics》 SCIE CAS CSCD 2008年第2期525-528,共4页
We propose an Ashkin-Teller-like model for elastic response of DNA molecule to external force and torque. The base-stacking interaction is described in a simple and uniform way. We obtain the phase diagram of dsDNA, a... We propose an Ashkin-Teller-like model for elastic response of DNA molecule to external force and torque. The base-stacking interaction is described in a simple and uniform way. We obtain the phase diagram of dsDNA, and in particular, the transition from 13 form to the S state induced by stretching and twisting. The elastic response of the ssDNA is presented also in a unified formalism. The close relation of dsDNA molecule structure with elastic response is shown clearly. The calculated folding angle of the dsDNA molecule is 59.2°. 展开更多
关键词 DSdna elastic property
下载PDF
Affinity chromatography-dependent selection (ACDS)of genomic DNA fragments bound specifically to bacterial synthesized Myc/Myn proteins
10
作者 SHI CAN PEI WANG +1 位作者 YONGJUN HU LIAN XU. (Oncogene Group, Laboratory of Molecular and Cellular Oncology, Shanghai Institute of Cell Biology, Chinese Academy of Sciences, 320 Yue-Yang Road, Shanghai 200031, China) 《Cell Research》 SCIE CAS CSCD 1995年第1期25-34,共10页
This paper describes an approach to seek for mouse c-Myc/Myn proteins-bound specific sequences among ge-nomic DNA. cDNA fragment of myn gene was obtained through RT-PCR technique from RNA of NIH3T3 cells. DNA fragment... This paper describes an approach to seek for mouse c-Myc/Myn proteins-bound specific sequences among ge-nomic DNA. cDNA fragment of myn gene was obtained through RT-PCR technique from RNA of NIH3T3 cells. DNA fragments encoding BR/HLH/LZ structure of Myc and Myn proteins were cloned in frame into pGEX-2T vec-tor respectively Fusion GST-Myc and GST-Myn synthe-sized in E.coli hosts showed affinity to CACGTG E-boxDNA and subsequently interacted with genomic fragments prepared through whole-genome-PCR. A PCR-assisted procedure which combines protein-DNA interaction and affinity chromatography was designed to enrich Myc/Myn bound DNA. At least two genomic DNA fragments ob- tained exhibit specifical binding capacity to Myc/Myn complex but not to GST alone. Significance of the work and of the technique itself as well as identification of the DNAs are discussed. 展开更多
关键词 Myc/Myn proteins ACDS genomic dna binding
下载PDF
Replication of M13 single-stranded DNA bearing a sitespecific ethenocytosine lesion by Escherichia coil cell extracts
11
作者 WANGGE PAULMDUNMAN MZAFRIHUMAYUN 《Cell Research》 SCIE CAS CSCD 1997年第1期1-12,共12页
Previous investigation on the mutagenic effects of 3,N4-Ethenocytosine (εC), a nonpairing DNA lesion,revealed the existence of a novel SOS-independent inducible mutagenic mechanism in E. coli termed UVM for UV modula... Previous investigation on the mutagenic effects of 3,N4-Ethenocytosine (εC), a nonpairing DNA lesion,revealed the existence of a novel SOS-independent inducible mutagenic mechanism in E. coli termed UVM for UV modulation of mutagenesis. To investigate whether UVM is mediated by an alteration of DNA replication, we have set up an in vitro replication system ill which phage M13 viral single-stranded DNA bearing a single site-specific (εC) residue is replicated by soluble protein extracts from E. coli cells. Replication products were analyzed by agarose gel electrophoresis and the frequency of translesion synthesis was determined by restriction endonuclease analyses. Our data indicate that DNA replication is strongly inhibited by εC, but that translesion DNA synthesis does occur in about 14% of the replicated DNA molecules. These results are very similar to those observed previously in vivo, and suggest that this experimental system may be suitable for evaluating alterations in DNA replication in UVM-induced cells. 展开更多
关键词 Ethenocytosine M13 in vitro replication cell extract
下载PDF
Mechanism of double-stranded supercoiled DNA cleavage induced by RNA N-glycosidase
12
作者 LIU Wang-yi WANG Hong-tao 《Journal of Life Sciences》 2009年第5期54-58,共5页
Plant RNA N-glycosidase specifically hydrolyzes the N-C glycosidic bond of a conserved adenosine in the sarcin/ricin domain of the largest RNA in ribosome, releasing an adenine base and thus inhibiting protein synthes... Plant RNA N-glycosidase specifically hydrolyzes the N-C glycosidic bond of a conserved adenosine in the sarcin/ricin domain of the largest RNA in ribosome, releasing an adenine base and thus inhibiting protein synthesis. This substrate specificity was challenged later by discovery that various RNA derivatives and DNAs, especially the double-stranded supercoiled DNA could be used as substrate by RNA N-glycosidase. Thus, it was argued whether the DNA-cleaving activity was an intrinsic feature of RNA N-glycosidase or it was contaminated by DNase. In this article, several lines of evidence are presented to show that RNA N-glycosidase can really release the adenine base from the double-stranded supercoi/ed DNA. It was proposed that the cleavage mechanism of supercoiled DNA was the phosphodiester bonds in enzymatically deadenylated regions of the supercoiled DNA would become fragile and liable to produce nicked or linear form owing to the existence of tension in the supercoiled DNA molecule, not direct result of enzymatic action on the phosphodiester bond. 展开更多
关键词 CINNAMOMIN releasing adenine RNA N-glycosidase supercoiled dna cleavage
下载PDF
芝田硫化叶菌Ssh7蛋白的进化保守性及DNA结合特性
13
作者 陈绪林 郭荣 +1 位作者 黄力 R.Hong 《中国科学(C辑)》 CSCD 北大核心 2002年第4期321-328,共8页
嗜酸热古菌——芝田硫化叶菌(Sulfolobus shibatae)合成大量7kD DNA结合蛋白Ssh7.Southern杂交结果显示,该菌基因组中含有两个编码Ssh7蛋白的基因,分别命名为ssh7a和ssh7b.对这两个基因进行了克隆、序列测定和在大肠杆菌中的表达.测... 嗜酸热古菌——芝田硫化叶菌(Sulfolobus shibatae)合成大量7kD DNA结合蛋白Ssh7.Southern杂交结果显示,该菌基因组中含有两个编码Ssh7蛋白的基因,分别命名为ssh7a和ssh7b.对这两个基因进行了克隆、序列测定和在大肠杆菌中的表达.测序结果表明,两个Ssh7多肽仅在3个氨基酸位置上有差异;此外,ssh7a和ssh7b的顺式调控序列也十分相似,提示存在着维持两个基因的序列和表达都不变的选择压力.杂交结果还显示,与芝田硫化叶菌一样,硫磺矿硫化叶菌(Sulfolobus solfataricus)基因组中也含有两个编码7 kD蛋白的基因.结合其他报道,这一结果提示编码7kD蛋白的基因可能在硫化叶菌种间分歧形成之前发生了基因复制.采用电泳迁移率改变试验(EMSA)分析了天然及重组Ssh7蛋白与双链DNA片段之间的相互作用.天然和重组蛋白的EMSA行为相似,说明Ssh7与DNA的相互作用既不受发生在天然蛋白赖氨酸残基上的甲基化的影响,也不受两种多肽异构体之间在序列上的差异的影响.在所采用的实验条件下,Ssh7与双链DNA片段结合时的结合位点大约为6.6bp,表观解离常数为(0.7~1.0)×10-4mol/L.此外,Ssh7与负超螺旋DNA的结合强于与线性和松弛DNA的结合. 展开更多
关键词 芝田硫化叶菌 Ssh7蛋白 进化保守性 dna结合特性 极端嗜热古菌 蛋白质dna-相互作用 dna结合蛋白
原文传递
The molecular characterization of maize B chromosome specific AFLPs 被引量:7
14
作者 ZHONG XIA QI, Hui ZENG, Xiu LAN LI, CHENG BIN CHEN, WEN QIN SONG, RUI YANG CHEN The College of Life Sciences, Nankai University, Tianjin 300071, China 《Cell Research》 SCIE CAS CSCD 2002年第1期63-68,共6页
The origin and evolution of B chromosomes could be explained by the specific DNA sequence on them. But the specific sequences known were quite limited. To investigate maize B chromosome sqicific DNA sequeces, maize ge... The origin and evolution of B chromosomes could be explained by the specific DNA sequence on them. But the specific sequences known were quite limited. To investigate maize B chromosome sqicific DNA sequeces, maize genomes with and without B chromosomes were analyzed by AFLP. Only 5 markers were found specific to genomes with B chromosomes among about 2000 AFLP markers. Southern hybridization and sequence analysis revealed that only the sequence of M8-2D was a B chromosome specific sequence. This sequence contained the telomeric repeat unit AGGGTTT conserved in plant chromosome telomeres. In addition, the sequence of M8-2D shared low homology to clones from maize chromosome 4 centromere as well. M8-2D were localized to B chromosome centrorneric and telomeric regions. 展开更多
关键词 MAIZE B chromosome AFLP FISH.
下载PDF
Development of a Real-Time PCR Method (Taqman) for Rapid Identification and Quantification of Prorocentrum donghaiense 被引量:1
15
作者 YUAN Jian MI Tiezhu +1 位作者 ZHEN Yu YU Zhigang 《Journal of Ocean University of China》 SCIE CAS 2012年第3期366-374,共9页
Prorocentrum donghaiense is a dinoflagellate that is widely distributed in the East China Sea and has become increasingly involved in Harmful Algal Blooms (HABs). Therefore, it is necessary to study this dinoflagellat... Prorocentrum donghaiense is a dinoflagellate that is widely distributed in the East China Sea and has become increasingly involved in Harmful Algal Blooms (HABs). Therefore, it is necessary to study this dinoflagellate to monitor HABs. In this study, 13 pairs of primers specific to P. donghaiense (within its internal transcribed spacer (ITS) regions) were designed for SYBR Green I real-time PCR. As the SYBR Green I real-time PCR could not identify P. donghaiense in a specific manner, a Taqman real-time PCR method was developed by designing a set of specific primers and a Taqman probe. A 10-fold serial dilution of recombinant plasmid containing ITS regions of P. donghaiense was prepared as standard samples and the standard curve was established. Additionally, we quantified the genomic DNA in P. donghaiense cells and utilized this DNA to prepare another 10-fold serial dilution of standard sample and accordingly set up the standard curve. The mathematic correlation between the cell number and its corresponding plasmid copy number was also established. In order to test the efficiency of the real-time PCR method, laboratory samples and P. donghaiense HAB field samples were employed for identification and quantitative analysis. As to laboratory samples, as few as 102 cells of P. donghaiense could be quantified precisely utilizing both centrifugation and filtration techniques. The quantification results from field samples by real-time PCR were highly similar to those by light microscopy. In conclusion, the real-time PCR could be applied to identify and quantify P. donghaiense in HABs. 展开更多
关键词 Prorocentrum donghaiense Harmful Algal Blooms (HABs) internal transcribed spacer (ITS) recombinant plasmid real-time PCR
下载PDF
Rapid,sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification 被引量:2
16
作者 高宏伟 李富花 +2 位作者 张晓军 王兵 相建海 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2010年第1期62-66,共5页
Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine... Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification (LAMP) assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase (empA) gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65℃ in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V, anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field. 展开更多
关键词 Loop-Mediated Isothermal Amplification (LAMP) detection assay empA gene Vibrio anguillarum
下载PDF
Panax ginseng-specific sequence characterized amplified region (SCAR) marker for testing medicinal products 被引量:1
17
作者 蒋秋桃 刘丽 +6 位作者 肖炳燚 李文莉 罗晖明 聂平 丁野 李洁 李文章 《Journal of Central South University》 SCIE EI CAS CSCD 2018年第5期1052-1062,共11页
To screen genetic polymorphisms of Panax ginseng, as well as those of Panax quinquefolium and Panax notoginseng, analysis of random amplified polymorphic DNA (RAPD) was performed using 120 random primers. Of the suc... To screen genetic polymorphisms of Panax ginseng, as well as those of Panax quinquefolium and Panax notoginseng, analysis of random amplified polymorphic DNA (RAPD) was performed using 120 random primers. Of the successful amplicons obtained, the Panax ginseng-specific RAPD marker C-12 was cloned into a TA vector and sequenced (Genl3ank access number KU553472). Based on the sequence analysis results, a pair of primers specific to C-12 was designed. Finally, a SCAR marker-based identification system for Panax ginseng was developed after optimization of the reaction conditions. Using this method, two positive bands were stably observed at 300 bp and 130 bp in 33 batches of Panax ginseng samples tested, while negative results were obtained for another 101 batches of samples, including Panax quinquefolium, Panax notoginseng, adulterants, and other medicinal herbs. Thus, we successfully developed a PCR-based method for rapid and effective identification of Panax ginseng, which can be effectively used for the protection and utilization of germplasm resources and identification of the origins of Panax ginseng samples. 展开更多
关键词 Panax ginseng random amplified polymorphic dna (RAPD) sequence characterized amplified regions(SCAR) molecular identification
下载PDF
Detecting Factor Ⅺ Deficiency in Holstein Cattle Using PCR Analysis
18
作者 张科 王占彬 王清义 《Agricultural Science & Technology》 CAS 2010年第5期109-111,共3页
[Objective] This study established a method to detect Factor Ⅺ by polymerase chain reaction analysis.[Method]A pair of primers was designed and synthesized according to sequences of FⅪ gene in Holstein calves,publis... [Objective] This study established a method to detect Factor Ⅺ by polymerase chain reaction analysis.[Method]A pair of primers was designed and synthesized according to sequences of FⅪ gene in Holstein calves,published in Genbank. Polymerase chain reaction was used to analyze FⅪ deficiency of 576 Holstein calves in Henan,and the result was verified by DNA sequencing. [Result] We detect 576 cows,which include two carriers and one F Ⅺ deficiency,and the result was consistent with the DNA sequencing. The frequency of the FⅪ mutant allele was 0.3%,the carrier was 0.3%,the prevalence was 0.2%.[Conclusion]A method detecting FⅪ by polymerase chain reaction analysis was established. This method is not only simple and convenient,but also has a high accuracy and low cost,which is more suitable for large-scale FⅪ investigation. 展开更多
关键词 Holstein cattle Factor deficiency PCR detection
下载PDF
Sequence-specific recognition of HIV-1 DNA based upon nicking-assisted strand displacement amplification and triplex DNA
19
作者 Manjun Zhang Ruimin Li +1 位作者 Jing Wang Liansheng Ling 《Science China Chemistry》 SCIE EI CAS CSCD 2017年第11期1468-1473,共6页
Nucleic acid amplification test is a reliable method for primary human immunodeficiency virus (HIV) infection diagnosis. Herein, a novel fluorescent method for sequence-specific recognition of DNA fragment of HIV-1 ... Nucleic acid amplification test is a reliable method for primary human immunodeficiency virus (HIV) infection diagnosis. Herein, a novel fluorescent method for sequence-specific recognition of DNA fragment of HIV-1 was established based upon nicking-assisted strand displacement amplification (SDA) and triplex DNA. In the presence of target dsDNA, nicking-assisted SDA process generated a lot of ssDNA, which hybridized with molecular beacon to produce signal. The fluorescence intensity was proportional to the concentration of target dsDNA within the range from 5 to 1000 pmol/L, with a detection limit of 1.4 pmol/L. Moreover, it successfully distinguished target dsDNA from the nucleic acid extractive of human blood. Thus this method has the merit of high sensitivity, and it is suitable for sequence-specific recognition of target dsDNA in complex matrices, which made it a potential application in diagnosis of acquired immunodeflciency syndrome (AIDS) in the future. 展开更多
关键词 HIV-1 dsdna nicking-assisted strand displacement amplification triplex dna Y-shaped structure fluorescence
原文传递
Enhanced destabilization of mismatched DNA using gold nanoparticles offers specificity without compromising sensitivity for nucleic acid analyses 被引量:1
20
作者 Abootaleb Sedighi Vicki Whitehall Paul C. H. Li 《Nano Research》 SCIE EI CAS CSCD 2015年第12期3922-3933,共12页
Here, we report a method that uses gold nanoparticles (AuNPs) to enhance the specificity of DNA hybridization without reducing its detection sensitivity. The conventional stringent wash method utilizes high-temperat... Here, we report a method that uses gold nanoparticles (AuNPs) to enhance the specificity of DNA hybridization without reducing its detection sensitivity. The conventional stringent wash method utilizes high-temperatureflow-salt conditions to enhance the specificity of DNA hybridization-based assays. This method creates a destabilizing environment for base pairing that affects specific and nonspecific duplexes. Therefore, specificity is achieved at the expense of signal intensity or sensitivity. However, in the proposed wash method, AuNPs predominantly destabilize nonspecific duplexes, offering specificity without compromising sensitivity. This AuNP wash technique has proven to be effective in detecting single nucleotide polymorphisms (SNPs) in genomic samples even at room temperature in a CD-like NanoBioArray (CD-NBA) chip. This method is also robust with sequence variation and is compatible with multiplex DNA analyses on microarrays. Thus, the AuNP wash method could potentially be useful for improving the accuracy of DNA hybridization results. 展开更多
关键词 dna hybridization gold nanoparticles single nucleotidepolymorphism SPECIFICITY CD-like NanoBioArray chip
原文传递
上一页 1 2 下一页 到第
使用帮助 返回顶部