光周敏感核不育水稻phyA基因的RFLP分析

本文用RFLP方法分析农垦58和光敏核不育水稻农垦58S以及其他一些水稻品种如IR36等，得到如下结果： 1．以第5染色体上8个RFLP标记(RG】3、RG】19、RG470、RG573、RG360、RG403、RG229、RG556)作探针对58和58S进行RFLP分析．没有发现多态性。 2． 58和58S光敏色素(phyA)基因内部（或附近）一些CpG岛的甲基化程度不同。 以IR36 phyA 基因的c DNA克隆作探针，用限制性内切酶EcoRV、Dra I、HindⅢ、Xba工、BglⅡ、Sca工、Msp工对58和58S进行RFLP分析，没有发现多态性，但用HpaII现了多态性。 3．根据IR36 ph yA序列，在5’一非编码区设计一对PCR引物，在IR36、58、58S等水稻品种中均扩出了一段DNA，此DNA片段大小在这几个不同品种中都相同。以此DNA片段（称为PCRI）作探针进行RFLP分析．发现在58S和IR36之间具有多态性，根据测出的58S的PCRI序列以及RFLP分析结果，推测58S和IR36之间的差异是由于插入或缺失引起的。

乌珠穆沁羊mtDNA的RFLP研究

用Apa I等7种识别6碱基的限制性内切酶研究了14只乌珠穆沁羊mtDNA的RFLP。结果表明,在所研究的个体中检测到37个酶切位点,17种限制性态型,其中BamH I和Bgl I表现出多态,Xho I无切点。17种限制性态型可归结为3种基因单倍型,mtDNA多态度π值为0.027%,表明乌珠穆沁羊mtDNA多态性比较贫乏。

绵羊、山羊和岩羊mtDNA的RFLP及其遗传分化研究

用15种识别6碱基的限制性内切酶ApaⅠ、BamHⅠ、BglⅠ、BglⅡ、ClaⅠ、DraⅠ、EcoRⅠ、EcoRⅤ、HaeⅡ、HindⅢ、KpnⅠ、PvuⅡ、PstⅠ、SacⅠ和SalⅠ对绵羊、山羊和岩羊mtDNA的限制性片断长度多态性进行了比较研究,以探讨其遗传分化关系。

Characterization of Sarcocystis species in domestic animals using a PCR-RFLP analysis of variation in the 18S rRNA gene: a cost-effective and simple technique for routine species identification

Thirteen restriction endonucleases were used to investigate nucleotide sequence variation in the 18S rRNA DNA of 88 individuals from ten Sarcocystis taxa collected as cysts from their intermediate hosts, swine, cattle and water buffalo. A DNA sequence of

A PCR-based RFLP analysis of Sarcocystis cruzi (Protozoa : Sarcocystidae) in Yunnan province, PR china, reveals the water buffalo (Bubalus bubalis) as a natural intermediate host

A polymerase chain reaction-based restriction fragment length polymorphism (RFLP) approach is used to examine Sarcocystis cruzi-like taxa from the atypical intermediate host, water buffalo, in Yunnan, People's Republic of China. The loci examined lie with

Investigation of polymorphism by PCR-RFLP method in Cobia fish Rachycentron canadum in the Persian Gulf and Oman Sea

Cobia is a native fish species in Iranian waters in the Persian Gulf and Sea of Oman and has a good internal and foreign market. This fish is a fast growing species and for this reason Iranian Fisheries is considering to go for it culture practices. To go for any utilization such as fishing from wild stocks or culture activities, needs a better understanding of its peculiarities and genetic characteristics of its natural resources. Therefore, this project was discribed and conducted. In this investigation, cuts 2 or 3 cm of fin tissue of  specimen of Cobia obtained from Sistan and Bluchestan, Hormozgan, Bushehr and Khuzestan water provinces, were collected. DNA was extracted by Phenol-chlorophorm method and produced PCR product in length of 1060 and 1450 base pair of two mitochondrial genes COI and NADH2. Using 13 cutting enzymes (4 enzymes were subscriber for both of genes), 205 base pair (from 2510 base pair, equal with %3.8 from gene regains) were directly investigated. But binding patterns of enzymatic digestion of PCR products of both COI and ND genes from electrophoresis were monomorph in all samples and no polymorphism was observed. This may be attributed to the unsuitable choice of COI and ND2 genes for showing of intra specific divergence. But in general non-existence of genetic diversity or noticeable decrease of that among individuals has been reported in regions were fish migration exist and they can freely move between two regions. Therefore, non-observation of polymorphism in the study area might be the case and indicates represents the area. On the other hand, some scientists believe that the distributions of populations in different regions are greatly affected by environmental and physical and ecological factors. Althoug Cobia is a migratory fish, but with regard to the fact that the environmental conditions are different (specially temperature and salinity) between east and west of Persian Gulf and Oman sea, there is a possibility that different genetic groups of this species exist in the regions. Of course It is clear that using more samples and enzymes from other genetically regions could produce better results. Since none of the two investigated genes didn’t show genetic divergence or polymorphism amongst the individuals of one region or between different regions, therefore, statistic analysis for estimating of haplotype diversity or nucleotide diversity and drawing of relationship tree among individuals using available softwares was not possible.

Genetic variation survey of intra-specific (Sepia pharaonis) in the Persian Gulf and the Oman Sea using PCR-RFLP

This research was conducted to identify Cuttlefish population (Sepia pharaonis) in The Persian Gulf and the Oman Sea using PCR-RFLP. Specimens were collected from )0 different stations. Bottom trawling method was used for sampling from different zones of the Persian Gulf and the Oman Sea, and finally specimens from S. Pharaonis were collected at each station . DNA was extracted by phenol—Coloroform method. One pair primer was designed based on 1As rRNA gene nucleotide sequences. The results obtained from 1 As rRNA gene RFLP, which was reproduced by PCR technique, were analyzed and utilized for study of diversity of the Cuttlefish population. PCR product with o pair base in length achieved for all specimens, which was subjected to enzymatic digestion by A restriction action enzymes: Alu I-Taq I-Mnl I-Rsa I-Hind III-Dra I-vu II and Hae II DNA bands patterns in all specimens digested by those enzymen showed similarity with no any polymorphism. From this result, it can be concluded that there is not any possibility to isolate different populations in the studied Cuttlefish species under exploitation of rRNA gene.

鲤鱼3个亚种线粒体DNA ND5/6区段的PCR-RFLP分析及亚种间分子标记的确立

从鲤鱼3个亚种(Cyprinus carpio carpio,Cyprinus carpio haematopterus和Cyprinus carpiorubrofuscus)中选出具代表性的品系共10个,运用PCR方法,扩增出2.4 kb的mtDNA ND5/6区段.共用9种限制性内切酶进行酶切分析,结果表明,每个亚种拥有一种单倍型,有4种限制性内切酶(Dde I,Hae III,Taq I和Mbo I)酶切后产生了能将3个亚种区分开来的诊断性限制性酶切位点.可利用其作为鉴定3个亚种的遗传标记和遗传育种

PCR-RFLP analysis of mitochondrial DNA ND5/6 region among 3 subspecies of common carp (Cyprinus carpio L.) and its application to genetic discrimination of subspecies

Mitochondrial DNA ND5/6 region was studied by PCR-RFLP analysis among ten representative strains belonging to three subspecies (Cyprinus carpio carpio, Cyprinus carpio haematopterus and Cyprinus carpio rubrofuscus) of common carp (Cyprinus carpio L.). A total of 2.4 kb fragment was amplified and subjected to restriction endonuclease analysis with nine restriction endonucleases subsequently. The results indicated that each subspecies owned one hyplotype and four restriction enzymes (Dde I, HaeIII, Taq I and Mbo I) produced diagnostic restriction sites which could be used for discriminating the three subspecies and as molecular genetic markers for assistant selective breeding of common carp.

铬污染土壤修复过程中土壤细菌群落多样性的RFLP分析

对污染土壤修复过程中土壤细菌群落多样性的变化进行研究。【方法】以淹水培养后的模拟铬污染土壤为供试材料,通过直接提取土壤中总细菌DNA,利用细菌专一引物克隆细菌16S rDNA片段,分别建立克隆文库。利用PCR-RFLP技术,分析比较了土壤淹水10 d(对照,S1)、添加Cr(Ⅵ)淹水10 d(S2)、添加Cr(Ⅵ)和Fe(OH)3淹水10 d(S3)及20 d(S4)4个处理中土壤细菌群落的变化。【结果】用专一引物克隆细菌16S rDNA片段,分别建立了克隆文库;用限制性内切酶RsaⅠ进行细菌16S rDNA PCR-RFLP分析,分别得到123,120,97和69个酶切类型,库容值分别为54.92%,55.43%,65.33%和76.60%;Shannon-Wiener指数、Gini指数、物种丰富度指数(dMa)和物种均匀度指数(Jgi)均表现为S1>S2>S3>S4,以上4个指数的变异系数分别为11.51%,1.84%,23.64%和1.55%;基于细菌多样性参数的聚类分析结果,将对照S1和添加Cr(Ⅵ)处理的S2归于一类,而2个添加Fe处理的土壤S3和S4聚为一类。【结论】经过10 d淹水处理,...

长白山四种赤杨丛枝菌根真菌侵染多样性的巢式PCR-RFLP分析

采集中国吉林长白山不同海拔的4种赤杨根须样本,利用巢式PCR-RFLP方法检测丛枝菌根真菌(AMF)对样品的侵染情况,PCR结果经限制性内切酶分析.结果表明,赤杨根内AMF存在丰富的基因多样性.AMF的侵染有从宿主混乱性向宿主专一性发展的趋势.东北赤杨AMF的宿主专一性水平最强,球囊霉属已成为东北赤杨的优势侵染类群;其余3种赤杨的AMF则出现宿主混乱现象.宿主因素比海拔因素对AMF侵染有更重要的影响.

高黎贡山旱冬瓜Frankia的IGSPCR-RFLP分析

在云南省高黎贡山自然保护区海拔 1310～ 2 4 0 0m的范围内 ,采集 30个旱冬瓜根瘤样品 ,直接从根瘤中提取FrankiaDNA ,对其nifD nifK基因间隔区 (intergenicspacer,IGS)和 16S 2 3SrDNAIGS进行PCR RFLP分析 .结果表明 ,nifD nifKIGS的PCR产物长度差异很大 ,经HaeⅢ和AfaⅠ双酶切后 ,得到 15种酶切带型 ,检测到多种基因型的菌株同时与同一株宿主植物共生 ;16S 2 3SrDNAIGS的PCR产物长度相似 ,酶切后亦区分出 15种酶切带型 .通过对两个基因间隔区的PCR RFLP联合分析 ,发现高黎贡山旱冬瓜Frankia存在 2 0种基因型

Species identification in salted products of red snappers by semi-nested PCR-RFLP based on the mitochondria 12S rRNA gene sequence

A molecular approach was developed to distinguish species of red snappers among commercial salted fish products. The specific fragments of the mitocliondrial 12S rRNA gene, which were about 450 bp, were obtained using the semi-nested polymerase chain reaction (semi-nested PCR). Subsequently, PCR amplicons were sequenced, aiming to select restriction endonucleases that generated species-specific restriction fragment length polymorphism (RFLP) profiles. Discrimination of red snappers Lutjanus sanguineus, L. erythopterus from L. argentintaculatus, L. malabarius and other morphologically similar fishes such as Lethrinus leutjanus and Pinjalo pinjalo was feasible by one restriction digestion reaction with three endonucleases Hae III, Sca I and SnaB I, however, for differentiation of L. sattguineus and L. erythopterus, another restriction digestion reaction with single restriction endonuclease Mae II was needed. The seminested PCR-RFLP was demonstrated to be reliable in species identification of salted fish products in this study. (c) 2005 Published by Elsevier Ltd.