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1.
大豆灰斑病菌生理小种的RAPD标记 总被引:8,自引:1,他引:7
运用随机扩增多态性DNA(RandomamplifiedpolymorphicDNA,RAPD)技术对发生于中国东北的大豆灰斑病菌(Cercosporidiumsojimum)的10个生理小种进行基因组DNA多态性分析,用13个10-核苷酸随机引物获得了111个RAPD标记,其中86.5%具有多态性,通过聚类分析确定了供试小种间的亲缘关系,试验证明,RAPD技术分析大豆灰斑病菌遗传传变异可提供大量 相似文献
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利用分子标记定位农垦58S的光敏核不育基因 总被引:17,自引:0,他引:17
对农垦58S(Oryzasativasp.japonica)/大黑矮生标记基因系FL2组合组建可育集团和不育集团,并以亲本为对照进行了RFLP、RAPD和双引物RAPD分析,结果第12染色体上的一个单拷贝标记G2140与光敏核不育基因连锁遗传,二者间的遗传图距为14.1cM(centimorgan)。在筛选过的1040个随机单引物和190个双引物中,仅引物OPAU10扩增出与光敏核不育基因连锁的1.5kbDNA片段,回收、克隆该DNA片段并制备探针,将其转换成共显性的RFLP标记并命名为OPAU101500。分离群体连锁分析表明该标记与标记G2140紧密连锁,将农垦58S的一对光敏核不育基因定位于第12染色体上。 相似文献
3.
黑子南瓜甘油-3-磷酸酰基转移酶基因的克隆及序列分析 总被引:6,自引:3,他引:3
依据国外报道的南瓜甘油-3-磷酸转酰酶(GPAT)基因的cDNA序列合成相应引物,用RT-PCR技术,成功地分离了黑子南瓜(Cucurbitaficifolia)GPAT基因的cDNA片段,并亚克隆到了pGEM-T载体系统的多克隆位点上,序列分析表明黑子南瓜GPAT基因的cDNA序列及递推的氨基酸序列与南瓜(Cucurbitamoschata)相比分别具有98%和965%的同源性。在1188bp中有22个核苷酸发生变化,导致13个氨基酸的改变 相似文献
4.
小麦胞质突变型雄性不育系85EA与其保持系线粒体DNA的比较研究 总被引:6,自引:1,他引:6
85EA是通过电子束辐照获得的胞质突变型小麦不育要用RFLP和RAPD技术对85EA及春保持系的线粒体DNA进行了比较研究。RFLP分析表明85EA线粒体基因组中coxⅡ基因的位置结构与保持系发生了变化;RAPD分析中引物OPD-15扩增产物在不育系和保持系间有明显差异,不育系的扩增产物比保持系多1条分子量为0.6kb的特展览 要带,用T-easy vector克隆该不育系特异条带并命名为OPD- 相似文献
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从端粒酶活性呈性阳的水生细胞株人肺SCP-A-1中分离了总RNA,以此为模板,结合RT-PCR技术和长模板PCR技术,用hTERT基因特异性引物扩增到一长约2.2kb的cDNA片段。将该片纯化后克隆到通用测序本T-easy vector上得到重组质粒。用测引物SP6和T7对该片段进行部分双向测序。经序列分析和同源比较推测该片段包含了hTERT基因的第3内含子。该结果提示了RT-PCR技术和长模板P 相似文献
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用pUC19质粒作载体,克隆了黄地老虎颗粒体病毒(Agrolissegetumgranulosisvirus,简称AsGV)DNAPstI-D.E.F.G.H.J.K.等7个片段。以[ ̄(32)P]-dCTP标记的油桐尺蠖核型多角体病毒(Buzurasuppressarianuclcarpolyhedrosisvirus简称BsNPV)多角体蛋白基因为探针,在37℃条件下对AsGV)颗粒体蛋白基因进行了定位,将其分别定位于BslⅡ-S或TPsTI-A或B和EciRI-A片段上。 相似文献
7.
鸡减蛋综合征病毒(EDSV—76)基因组E1区结构特点分析 总被引:1,自引:0,他引:1
EDSV-76病毒中国株AA-2经常规方法提取其病毒DNA后,建立了限制性内切酶PstI水解片段的全基因文库。对其中PstI-G片段和PstI-A片段的正反链进行序列测定,获得EDSV E1区(0-8.8m.u)的核苷酸序列。经分析,EDSV E1区具有与其他腺病毒E1区类似的结构。以大于60个氨基酸残基为标准,EDSV E1区共有7个开放读码框架(ORF),其中R1、R2、ElbsT和E1b1T 相似文献
8.
DNA分子标记在柑桔中的应用 总被引:3,自引:1,他引:2
DNA分子标记是最为理想的遗传标记 ,依其多态性检出所用的分子生物学技术 ,大致可分为Southem杂交技术为核心的分子标记和PCR技术为核心的分子标记。前者的代表性技术有RELP(restrictionfragmentlengthpolymorphism)和DNA指纹技术 (DNAfingerprintingtechniques)。后者的代表性技术有RAPD(randomamplifiedpolymorphicDNA)、SCAR(sequencecharac teristicamplifiedregion)… 相似文献
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RAPD技术及其在微生物学方面的应用 总被引:4,自引:0,他引:4
198 0年 ,Botsein提出DNA限制性片段长度多态性 (RFLP)可以作为遗传标记 ,从此开创了直接应用DNA多态的新阶段。 80年代后 ,DNA多聚酶链式反应 (PCR)的发展 ,使直接扩增DNA的多态性成为可能 ,并在此基础上产生了许多种新型分子标记 ,诸如扩增片段多态性 (ALFR)、串联重复序列(VNTR)、单链构型多态性 (PCR SSCP)、序列特异扩增区域 (SCAR)、随机扩增多态性DNA(RAPD)等。而RAPD是较为突出的一种。RAPD是由Williams和Welsh在 1 990年各自独立发现的一种DNA多态检… 相似文献
11.
葡萄感霜霉病基因的分子标记(英文) 总被引:4,自引:0,他引:4
在葡萄抗病育种中 ,幼苗期排除感霜霉病的后代具有特别重要的意义 .用 BSA,RAPD和SCAR方法研究了葡萄感霜霉病基因的分子标记 .分析了两个种间杂交组合 [毛葡萄 (抗病 )×欧洲葡萄 (感病 ) ]88- 1 1 0和 88- 84与 88- 1 1 0的 F1代自交或互交所得的 3个 F2 代 ,以及欧洲葡萄品种和中国野生葡萄种 .共筛选了 2 80个随机引物 .引物 OPO1 0产生了一个 RAPD标记 OPO1 0 - 80 0与葡萄感霜霉病主效基因紧密联锁 .将该 DNA片段克隆并测序 .OPO1 0 - 80 0的实际长度为 835bp,所以 OPO1 0 - 80 0应为 OPO1 0 - 835.据其两端序列 ,设计了一对长度为 2 6bp和 2 8bp的特异引物分别扩增上述试材 ,获得了与该 RAPD标记相同大小的一条带 ,将 RAPD标记转化为 SCAR标记SCO1 0 - 835.并证实了此 SCAR标记的通用性 ,该 SCAR标记可用于葡萄抗病育种中杂种后代对霜霉病的抗病与感病性鉴定 . 相似文献
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Jung Sou Yeo Ji Sun Lee Chang Hee Lee Young Ja Jung Doo Hyun Nam 《Biotechnology and Bioprocess Engineering》2000,5(1):23-26
In order to develop the specific genetic marker for Korean native cattle (Hanwoo), randomly amplified polymorphic DNA (RAPD)
analysis of 6 different cattle breeds was attempted by using 38 decamer primers. In comparison of RAPD patterns, two distinctive
DNA bands specific for Hanwoo were detected. One was 296 bp of DNA fragment found to be specific only for female Hanwoo when
primer GTCCACACGG was employed. In individual analysis of this RAPD marker was observed only in female individuals with the
possibility of 85.3%. The other was 521 bp of RAPD marker amplified using TCGGCGATAG and AGCCAGCGAA primers, which showed
83.0% of genetic frequency in 85 male and 68 female individuals tested. Nucleotide sequencing of these genetic markers revealed
that 296 bp marker has a short microsatellite-like sequence, ACCACCACAC, and a tandem repeat sequence of microsatellite GAAAAATG
in the determined sequence. Two distinctive tandem repeats of microsatellite sequences, AAC and GAAGA, were also appeared
in 521 bp DNA marker. In BLAST search, any gene having high homology with these markers was not found 相似文献
14.
黄瓜霜霉病抗病基因的RAPD及SCAR标记 总被引:3,自引:0,他引:3
以感霜霉病黄瓜L18-10-2和抗霜霉病黄瓜129为亲本构建F2代分离群体,以F3代植株霜霉病抗性鉴定表示F2代各单株抗病性并得以区分各单株杂合或纯合感病性,采用RAPD技术和转SCAR的方法筛选黄瓜抗霜霉病基因分子标记.结果显示,在318条RAPD引物中有18条引物表现出两亲本间多态性,其中引物P18的SB-SP18561扩增片段与霜霉病抗病基因之间紧密连锁,根据交换率和Kosambi函数公式计算其遗传距离为7.85 cM.回收SBSP18561片段并克隆和测序,其准确长度为561 bp.将该RAPD标记转换为SCAR标记,长度为494 bp,命名为SSBSP18494. 相似文献
15.
D. Ladhalakshmi A. Vijayasamundeeswari V. Paranidharan R. Samiyappan R. Velazhahan 《World journal of microbiology & biotechnology》2009,25(12):2129-2135
The fungus Peronosclerospora sorghi [Weston and Uppal (Shaw)] infects both sorghum and maize and incites downy mildew disease. Pathogenic and molecular variability
among isolates of P. sorghi from sorghum and maize has been reported. In the present study we developed a DNA sequence characterized amplified region
(SCAR) marker for identification of isolates of P. sorghi from maize by using polymerase chain reaction (PCR). The random amplified polymorphic DNA (RAPD) primer OPB15 consistently
amplified a 1,000 base pairs (bp) product in PCR only from DNA of P. sorghi isolates from maize and not from isolates of sorghum. The PCR-amplified 1,000-bp product was cloned and sequenced. The sequence
of the SCAR marker was used for designing specific primers for identification of maize isolates of P. sorghi. The SCAR primers amplified a 800 bp fragment only from genomic DNA of maize isolates of P. sorghi. The SCAR primers developed in this study are highly specific and reproducible, and proved to be powerful tool for identification
of P. sorghi isolates from maize. 相似文献
16.
M A Gedil M B Slabaugh S Berry R Johnson R Michelmore J Miller T Gulya S J Knapp 《Génome》2001,44(2):205-212
Disease resistance gene candidates (RGCs) belonging to the nucleotide-binding site (NBS) superfamily have been cloned from numerous crop plants using highly conserved DNA sequence motifs. The aims of this research were to (i) isolate genomic DNA clones for RGCs in cultivated sunflower (Helianthus annuus L.) and (ii) map RGC markers and Pl1, a gene for resistance to downy mildew (Plasmopara halstedii (Farl.) Berl. & de Toni) race 1. Degenerate oligonucleotide primers targeted to conserved NBS DNA sequence motifs were used to amplify RGC fragments from sunflower genomic DNA. PCR products were cloned, sequenced, and assigned to 11 groups. RFLP analyses mapped six RGC loci to three linkage groups. One of the RGCs (Ha-4W2) was linked to Pl1, a downy mildew resistance gene. A cleaved amplified polymorphic sequence (CAPS) marker was developed for Ha-4W2 using gene-specific oligonucleotide primers. Downy mildew susceptible lines (HA89 and HA372) lacked a 276-bp Tsp5091 restriction fragment that was present in downy mildew resistant lines (HA370, 335, 336, 337, 338, and 339). HA370 x HA372 F2 progeny were genotyped for the Ha-4W2 CAPS marker and phenotyped for resistance to downy mildew race 1. The CAPS marker was linked to but did not completely cosegregate with Pl1 on linkage group 8. Ha-4W2 was found to comprise a gene family with at least five members. Although genetic markers for Ha-4W2 have utility for marker-assisted selection, the RGC detected by the CAPS marker has been ruled out as a candidate gene for Pl1. Three of the RGC probes were monomorphic between HA370 and HA372 and still need to be mapped and screened for linkage to disease resistance loci. 相似文献
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采用282个随机引物对药用野生稻1665和栽培稻桂99远缘杂交的抗褐飞虱近等基因系B3F4分离群体的不抗池DNA和抗池DNA进行了特异性RAPD标记筛选,从中筛选到一个具有明显的特异性扩增带谱的RAPD标记S1159,序列分析表明,S1159序列长度为1408bp,与基因库中已报道的水稻第四号染色体的BAC克隆(编号OSJNBa0070O11)序列(67114-69100)有51.86%的同源性。为了提高所找到的RAPD标记S1159在应用上的稳定性,将RAPD标记转化为SCAR标记检测近等基因系群体,结果表明与RAPD标记结果一致,说明该研究得到的RAPD标记具有较好的稳定性和重复性,为进一步的研究打下了良好的基础。 相似文献
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Abstract A method is described for developing a sheep‐ vs. goat‐specific DNA marker using sequence characterized amplified regions (SCARs) derived from a random amplified polymorphic DNA (RAPD) marker from sheep DNA samples. A sheep 645 bp DNA fragment that was absent in goat DNA was identified by analyzing pools of sheep and goat DNA with RAPD primers. This fragment was cloned and partially sequenced to design extended, strand‐specific 24‐mer oligonucleotide primers. Each primer contained the original 10 bases of the RAPD primer and the following 14 internal bases. The pair of primers resulted in the amplification of a single band of 645 bp when used to amplify sheep DNA, and in no amplification when used to amplify goat DNA. These SCAR primers successfully amplified the equivalent of DNA from one nucleated sheep cell in a sample of 5000 nucleated goat cells. This level of sensitivity is especially desirable for research involving the detection of interspecific chimerism. 相似文献
20.
Kim S Song YH Lee JY Choi SR Dhandapani V Jang CS Lim YP Han T 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2011,123(7):1183-1192
Inheritance of resistance to downy mildew (Hyaloperonospora parasitica) in Chinese cabbage (Brassica rapa ssp. pekinensis) was studied using inbred parental lines RS1 and SS1 that display strong resistance and severe susceptibility, respectively. F(1), F(2), and BC(1)F(1) populations were evaluated for their responses to downy mildew infection. Resistance to downy mildew was conditioned by a single dominant locus designated BrRHP1. A random amplified polymorphic DNA (RAPD) marker linked to BrRHP1 was identified using bulked segregant analysis and two molecular markers designated BrPERK15A and BrPERK15B were developed. BrPERK15B was polymorphic between the parental lines used to construct the reference linkage map of B. rapa, allowing the mapping of the BrRHP1 locus to the A1 linkage group. Using bacterial artificial chromosome clone sequences anchored to the A1 linkage group, six simple polymerase chain reaction (PCR) markers were developed for use in marker-assisted breeding of downy mildew resistance in Chinese cabbage. Four simple PCR markers flanking the BrRHP1 locus were shown to be collinear with the long-arm region of Arabidopsis chromosome 3. The two closely linked flanking markers delimit the BrRHP1 locus within a 2.2-Mb interval of this Arabidopsis syntenic region. 相似文献