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1.
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   

2.
杂交水稻及其“三系”线粒体DNA的AP—PCR指纹图谱   总被引:19,自引:1,他引:18  
为了研究水稻(Oryza sativa L.)细胞质雄性不育(CMS)与线粒体基因组的关系,应用AP-PCR 分析,用7 个任意单引物对6 种水稻品系线粒体DNA 进行了扩增。水稻线粒体DNA 的AP-PCR 产物可分为三种类型:(1)所有供试品系均能扩增的片段,它们代表了线粒体DNA 在进化上的保守性序列。有4 个引物检测到这类片段。(2)2 个以上水稻品系共同出现而在全部供试材料间存在差异的扩增片段,这类片段是检测水稻线粒体DNA多态性的主要来源。(3)一种细胞质类型所特有的扩增片段,从引物R2 和V5 的扩增产物中发现了这类片段,它们可能与CMS有关联。另外,WA型不育系珍汕97A 与其杂种之间在6 个引物的扩增图谱上均存在不同程度的差异,说明两者的线粒体DNA序列结构可能存在某种差别  相似文献   

3.
孙易  宋文芹 《遗传学报》2000,27(3):210-218
利用RAPD(随机扩增多态DNA)和AFLP(扩增片段长度多态性)技术对不同种及种群卤虫的关系进行分析。101个随机引物对4种卤虫A.franciscana、A.urmiana、A.sinica和A.parthenogenetica基因组DNA进行扩增,平均每个种获得751条带,其中458条带为多态性标记,每个引物提供平均7.4个标记信息,聚类结果表明A.sinica是不同于其他旧大陆两性生殖卤虫  相似文献   

4.
RAPD在植物育种上的应用及其技术校正   总被引:1,自引:0,他引:1  
RAPD(RandomAmplifiedPolymorphicDNA) ,是在PCR的基础上发展起来的一种DNA多态性检测技术 ,由Willams[1] 和Welsh[2 ] 于 1990年建立 ,原理是以一系列不同的 ,少数碱基组成的随机核甘酸序列为引物 ,对样本基因组DNA进行PCR扩增 ,每个RAPD片段的产生要求在可增范围内存在与引物匹配的反向互补序列。引物结合位点DNA序列的改变以及两扩增位点之间碱基的缺失插入或转换都能导致扩增片段的数目和长度的差异 ,经PAGE或琼脂糖凝胶电泳分离和EB染色来检测DNA片段的多态…  相似文献   

5.
以菠菜(Spinacia oleracea L.)为材料,取幼叶分离mRNA,反转录合成cDNA,以cDNA第一链为模板,通过PCR扩增,获得菠菜磷酸丙糖转移蛋白(Triose phosphate translocator,TPT)cDHA目的片段。对其进行序列分析,结果表明,分离的目的片段核苷酸序列与文献报道相比同源率为99.9%,只不1个碱基发生改变。将得到的菠菜tpt cDNA与CaMV35  相似文献   

6.
应用随机扩增多态DNA(RandomamplifiedpolymorphicDNAs,RAPD)技术,对普通小麦细胞质雄性不育系(A)及其保持系(B)线粒体基因组(mitochondrialgenome)的指纹图谱进行了分析。共使用引物41个,其中20个引物得到了扩增产物,部分引物扩增结果表现多态性。说明线粒体DNA(mitochondrialDNA,mtDNA)序列在不育系和保持系之间存在差异,进一步明确了小麦细胞质雄性不育(cytoplasmicmalesterile,CMS)与mtDNA的关系。  相似文献   

7.
应用随机扩增多态DNA(Random amplified polymorphic DNAs,RAPD)技术,对普通小麦细胞质雄性不育系(A)及其保持系(B)线粒体基因组(mitochondrial genome)的指纹图谱进行了分析。共使用引物41个,其中20个引物得到了扩增产物,部分引物扩增结果表现多态性。说明线粒体DNA(mitochondrial DAN,mt,DNA)序列在不育系和保持系之  相似文献   

8.
利用PCR技术分别以亲本杉木(Cunninghamialanceolata(Lamb.)Hook.)、柳杉(CryptomeriafortuneiHooibrenk)和杉木×柳杉杂种的总DNA为模板,扩增了叶绿体trnLtrnF和线粒体CoxⅢ基因片段,比较了这些扩增片段的限制性内切酶AluⅠ,DdeⅠ,HinfⅠ,MseⅠ和RsaⅠ的酶切片段多态性,结果表明:F1代的叶绿体DNA为母系遗传,而线粒体DNA为父系遗传。杉木线粒体DNA父系遗传方式与杉科其他植物一致,而叶绿体DNA母系遗传则为在松柏类植物中首次发现。  相似文献   

9.
王晓沁  李元 《遗传学报》1999,26(4):288-294
以分离提取的HeLa细胞总RNA为模板,通过RT-PCR反应扩增得到了1017bp的人可溶性白细胞介素6受体(sIL-6R)cDNA片段,将扩增片段克隆到pUC19质粒中进行序列测定。结果证明该片的序列与文献报道的sIL-6RcDNA的序列完全一致,将sIL-6RcDNA与链霉素信号肽melCl的编码序列融合后得到的融合基因mel/sIL-6R克隆到链霉菌质粒pLJ459中,构建成重组表达质粒pL  相似文献   

10.
用聚合酶链式反应(PCR)将豆科黄芪亚族7属9种及甘草亚族1属1种植物叶绿体基因组中ndhF和psbA基因中一段约3.1kb的DNA扩增出来,并摸索出一最佳的PCR条件,使得此条带得以特异性扩增,可直接用于限制性内切酶水解。通过对此扩增片段的限制性片段长度多态性(RFLP)的初步分析,认为利用PCR扩增出的DNA进行RFLP分析来探讨植物的系统进化问题在中国现行条件下大有潜力,其方法简单易行,结果  相似文献   

11.
测定了东海带鱼(Trichiurus lepturus Linnaeus)三个群体(命名为A: 122°32′E 29°55′N、B: 123°30′E 26°75N; C: 124°24′E 27°26′N)72个个体的线粒体DNA16S rRNA和COⅠ基因序列,通过单基因序列和联合序列分析, 研究了东海带鱼群体的遗传变异情况。分别得到1130 bp的COⅠ基因序列片段和554 bp的16S rRNA序列片段, 其中COⅠ基因片段的T、C、A、G含量分别为29.0%、28.9%、24.4%和17.7%; 16S rRNA基因片段的T、C、A、G含量分别为22.7%、27.6%、28.0%和21.7%。基于线粒体16S rRNA、COⅠ和16S+COⅠ基因序列分析, 72个个体中分别确定43个, 8个和49个单倍型, 存在单倍型共享现象。群体的单倍型多样性指数为0.9766—0.9992, 显示了群体内的单倍型较为丰富。3个群体间各序列平均核苷酸差异数(K)在5.111—9.024和0.0045—0.0076, 核苷酸多样性指数(π)为0.0048—0.0084, 显示不同带鱼群体遗传多态性丰富。使用邻接法构建的分子进化树揭示同一群体内大部分个体聚在一起。分析结果表明, 群体A遗传背景比群体B、群体C较为丰富, 群体内部个体差异大于群体间差异, 群体间基因交流频率较高, 遗传分化不明显, 初步判定东海带鱼3个群体的遗传多样性偏低。  相似文献   

12.
长吻(鱼危)(Leiocassis longirostris)是中国土著珍稀鱼类。近年来, 由于江河水利工程、环境污染及人类生产活动已经对江河的渔业资源造成了难以逆转的破坏, 长吻(鱼危)的渔业资源已逐渐枯竭。目前, 长吻(鱼危)在四川、广东等地实现适度规模养殖。以四川眉山、湖北石首和安徽淮南3个人工养殖长吻(鱼危)群体及4个长江野生长吻(鱼危)群体(重庆段、武汉段、安庆段和南京段)为实验材料, 利用线粒体DNA (mtDNA)控制区序列作为分子标记对135个个体的遗传结构进行了分析。结果表明, 在790 bp 的同源序列中, 长吻(鱼危) 3个养殖种群共检测到变异位点27个, 占全部序列的3.42%, 66个个体共检测到18种单倍型; 在野生群体中, 69个个体共检测到35个变异位点和36个单倍型, 长吻(鱼危)野生群体平均单倍型多样性和平均核苷酸多样性(Hd=0.9736±0.0070, Pi=0.0087±0.0015)高于长吻(鱼危)养殖群体(Hd=0.8867±0.0013, Pi=0.0056±0.0013); 群体间的遗传分化水平较低(Fst值为0.0014—0.1125)。采用邻接法(NJ法)和统计简约原理对所有单倍型进行系统发育树和统计简约网状图的构建, 结果表明: 各群体内的个体均不能分别构成独立的分支, 而是相互交叉聚在一起。分析结果表明, 长吻(鱼危)养殖群体与野生群体之间的基因交流充分, 未出现遗传分化, 但相对长吻(鱼危)野生群体, 长吻(鱼危)养殖种群多态性偏低。  相似文献   

13.
Guo E  Liu Y  Cui Z  Li X  Cheng Y  Wu X 《Molecular biology reports》2012,39(2):1453-1463
Genetic variation and population structure in Portunus trituberculatus along the coast of China were revealed according to 617 bp of mitochondrial DNA control region. 90 polymorphic sites defined 53 distinct haplotypes, showing a moderately high diversity among 72 individuals sampled from eight localities. Neighbor-joining tree, statistics analyses of gene flow and genetic differentiation index indicated two populations from Beihai and Laizhou had differentiated. The population from Yingkou, Dandong, Laizhou and Beihai had smaller genetic diversity compared to that from Ningbo, Lianyungang, Qingdao and Japan according to the genetic distance. And mantel test showed significant positive correlation between genetic distance and geographic distance for P. trituberculatus. TCS parsimony network suggested that all the animals sampled were probably the result of recent divergence from a common ancestral haplotype but for Laizhou population. Moreover, the haplotype distribution appeared to correlate with a recent colonization followed by localized genetic differentiation. Mismatch distribution results suggested that Ningbo, Yingkou, Qingdao, Lianyungang and Japan populations, particularly Dandong population had experienced a sudden demographic or spatial expansion. The Pleistocene glaciations might contribute to this process.  相似文献   

14.
基于线粒体Cyt b基因的龙头鱼群体遗传结构分析   总被引:1,自引:0,他引:1  
在我国黄海、东海和南海海岸带上, 选取青岛、南通、舟山、三门、宁德、泉州和湛江共7个群体164尾龙头鱼(Harpadon nehereus)为研究对象, 通过PCR扩增共获得长度为1112 bp的线粒体DNA细胞色素b (Cyt b)基因序列, 共检测到32个变异位点, 其中单变异位点27个, 简约信息位点1个。164条序列定义了29个单倍型, 平均单倍型多样性(Hd)和核苷酸多样性(π)分别为0.3026±0.0479和0.000371±0.000379, 其中泉州群体的单倍型多样性和核苷酸多样性均最低。分析比较了不同龙头鱼群体间遗传变异情况, 发现群体间平均遗传距离为0.00035, 遗传分化指数FST值均小于0.05, 群体间没有发生明显的遗传分化。AMOVA分析结果显示龙头鱼群体遗传差异主要来源于群体内个体间的变异。中性检验的Tajima’s D和Fu’s Fs统计值均为负值且显著偏离中性, 核苷酸不配对分布图呈现明显的单峰分布, 表明龙头鱼历史上经历了群体扩张事件, 参考已知海洋鱼类Cyt b基因2%每百万年的进化速率, 估算群体扩张发生的时间大约在0.08—0.32百万年前的第四纪更新世中晚期。  相似文献   

15.
In order to describe the genetic diversity of five geographical populations of cuttlefish (Sepiella japonica) along with Chinese coast and determined their phylogenetic relationship, partial mitochondrial COI gene (681bp in length) was amplified from 96 individuals collected from these populations and sequenced. The 5 populations of cuttlefish inhabit Yellow Sea, East China Sea and South China Sea. Out of 22 polymorphic nucleotides identified, 8 were represented by a single sequence, 12 were parsimony informative, which defined 22 haplotypes. Haplotype and nucleotide diversity were low among populations. Of 22 haplotypes, 15 appeared only in a single population, 6 appeared in 2 or 3 populations and 1 was shared by all populations. The COI gene was monomorphic in Qingdao population. The haplotypes identified clustered into 2 clades, each covered individuals from 5 populations each. Pairwise FST were not proportional to the geographical distances. Among the 5 populations, relatively high level of genetic diversity was found in Ningde population which was recommended to be the best choice of germplasm resources for artificial releasing project.  相似文献   

16.
To analyze the genetic diversity and genetic structure of cultured populations of Mytilus galloprovincialis, 44 individuals were sampled from three localities, Yantai, Rushan of Shandong province and Dongji of Zhejiang province. Sequence analysis of ribosomal (18S rRNA gene) and mitochondrial DNA (COIII gene) revealed 23 haplotypes of 18S rRNA and 30 haplotypes of COIII. Haplotype diversities (Hd), nucleotide diversities (Pi) and average nucleotide differences (K) were 0.712 (18S rRNA) and 0.946 (COIII), 0.0044 (18S rRNA) and 0.0207 (COIII), and 0.703 (18S rRNA) and 15.316 (COIII), respectively. Fixation indices (ΦST) of the three cultured populations showed no genetic divergence between Yantai and Rushan populations but significant genetic divergence between Dongji and the other two populations. In the present study, we found no evidence for decline in genetic polymorphism of cultural M. galloprovincialis populations. The findings of the present study will be useful in the management and conservation of M. galloprovincialis resources.  相似文献   

17.
By using six polymorphic microsatellite loci, the population genetic diversity and structure of 120 individuals of Portunus trituberculatus were examined in five wild populations representing different geographical and ecological ranges along the coast of China. Middle level of genetic diversity of P. trituberculatus was revealed by observed heterozygosity, allele number and Shannon information index. Pairwise population differentiations were found between all pairs of populations except Beihai (BH) and Qingdao (QD) (P < 0.05). The greatest differentiation was detected between Yingkou (YK) and QD populations. It was inconsistent with the result in previous study using mitochondrial control region, which showed no differentiation between QD and YK population. Recent bottleneck was identified in Ningbo (NB) population under TPM and IAM models, as well as Dandong (DD) and QD populations under IAM model. Additionally, compared with YK, QD, DD and BH populations, lower historical effective population size with lower genetic diversity were detected in NB population, which called for urgent assessment of the ecological and economic potential of NB fisheries.  相似文献   

18.
Abstract.  Variation in the mtDNA 16S ribosomal RNA gene in populations of Triatoma infestans (Klug) was surveyed. DNA sequence comparisons yielded 18 haplotypes among 130 individuals from 16 localities that represent a large proportion of the range of T. infestans in Argentina. The most common genotype in all populations was found in 76.9% of individuals and two other haplotypes were shared among different populations. The remaining 15 haplotypes were present exclusively in one of the populations, suggesting currently low levels of genetic exchange. Analysis of mtDNA 16S sequences uncovered substantial genetic variation among T. infestans populations. Haplotype and nucleotide diversities varied among populations, from 0% to 0.84% and 0% to 0.29%, respectively. It appears that this locus has a low mutation rate. Uncorrected pairwise differences of T. infestans haplotypes ranged from 0% to 1.2%. The molecular phylogeny supported the monophyly of T. infestans haplotypes and clustered two different pairs of haplotypes with a moderate degree of bootstrap support (∼ 60%). Mitochondrial DNA phylogeographic differentiation was not evident, suggesting a recent rapid spread of the species. Analysis of molecular variance showed hierarchical structure in the data. Considerably less variation was found among T. infestans populations from the northwest and northeast regions than among those belonging to the central area. Such a lack of variation may be indicative of one or more past population bottlenecks.  相似文献   

19.
The cocktail shrimp Trachypenaeus curvirostris is an ecologically and economically important shrimp species in the Yellow Sea and East China Sea. However, there is no information about its population genetic structure. The population genetic structure and level of gene flow of T. curvirostris from the Yellow Sea and the East China Sea were studied with a 658-bp segment of mtDNA COI gene. In total, 85 individuals were collected from five locations and 13 haplotypes were obtained. The genetic variation of COI gene in five populations was moderate, giving an overall haplotype diversity of 0.6888 ± 0.0432 and nucleotide diversity of 0.0069 ± 0.0038. Conflicting to our expectation, significant genetic differentiation was detected in this species. The result revealed two genetically divergent lineages, displaying clear different geographical distributions in the studied area. The significant genetic differentiation between the Yellow Sea and East China Sea populations might be caused by the Yangtze River outflow. Mismatch distribution revealed that T. curvirostris had undergone population range expansion, possibly before 103,400–109,700 years ago in the last interglaciation, rejecting the sudden demographic expansion.  相似文献   

20.
The Xianbei existed as a remarkable nomadic tribe in northeastern China for three dynasties: the Han, Jin, and Northern-Southern dynasties (206 BC to 581 AD) in Chinese history. A very important subtribe of the Xianbei is the Murong Xianbei. To investigate the genetic structure of the Murong Xianbei population and to address its genetic relationships with other nomadic tribes at a molecular level, we analyzed the control region sequences and coding-region single nucleotide polymorphism markers of mtDNA from the remains of the Lamadong cemetery of the Three-Yan Culture of the Murong Xianbei population, which is dated to 1,600-1,700 years ago. By combining polymorphisms of the control region with those from the code region, we assigned 17 individuals to haplogroups B, C, D, F, G2a, Z, M, and J1b1. The frequencies of these haplogroups were compared with those of Asian populations and a multidimensional scaling graph was constructed to investigate relationships with other Asian populations. The results indicate that the genetic structure of the Lamadong population is very intricate; it has haplogroups prevalent in both the Eastern Asian and the Siberian populations, showing more affinity with the Eastern Asian populations. The present study also shows that the ancient nomadic tribes of Huns, Tuoba Xianbei, and Murong Xianbei have different maternal genetic structures and that there could have been some genetic exchange among them.  相似文献   

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