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1.
Abundance, polymorphism and genetic mapping of microsatellites in rice   总被引:71,自引:0,他引:71  
Dinucleotide microsatellites have been characterized and used as genetic markers in rice. Screening of a rice genomic library with poly(dG-dA)·(dC-dT) and poly(dG-dT)·(dC-dA) probes indicated that (GA)n repeats occurred, on average, once every 225 kb and (GT)n repeats once every 480 kb. DNA sequencing of ten randomly selected microsatellites indicated that the numbers of repeats ranged from 12 to 34 and that the patterns of microsatellites in rice were similar to those of humans and other mammals. Primers to these microsatellite loci as well as to four published microsatellite-containing sequences have been designed and degrees of polymorphism has been examined with 20 rice accessions. Multiple alleles, ranging from 5 to 11, have been observed at all the microsatellite loci in 20 rice accessions. Alleles specific to two cultivated subspecies, indica and japonica, were found in some microsatellite loci. Heterozygosity values of all the microsatellite markers were significantly higher than those of RFLP markers, based upon a parallel comparison. Ten microsatellite loci have been genetically mapped to four rice chromosomes. The genomic distribution of microsatellites appears to be random in rice.  相似文献
2.
Analysis of SSRs derived from grape ESTs   总被引:70,自引:0,他引:70  
One hundred and twenty four microsatellites were isolated from analysis of 5000 Vitis expressed sequence tags (ESTs). A diversity of dinucleotide and trinucleotide simple sequence repeat (SSR) motifs were present. Primers were designed for 16 of these SSRs and they were tested on seven accessions. Ten of the sixteen primer pairs resulted in PCR products of the expected size. All ten functional primers were polymorphic across the accessions studied. Polymorphisms were evident at the level of cultivars, Vitis species, and between related genera. SSRs that were from the 3′ untranslated region (3′UTR) were most polymorphic at the cultivar level, the 5′ untranslated region (5′ UTR) SSRs were most polymorphic between cultivars and species, and those SSRs within coding sequence were most polymorphic between species and genera. These results show that EST-derived SSRs in Vitis are useful as they are polymorphic and highly transferable. With EST SSRs being applicable to studies at several taxonomic levels, the large number of SSRs (approximately 1000) that will be available from an expanded EST database of 45 000 will have many potential applications in mapping and identity research. Received: 4 June 1999 / Accepted: 21 September 1999  相似文献
3.
鲤鱼微卫星分子标记的筛选   总被引:66,自引:3,他引:63       下载免费PDF全文
微卫星 (microsatellite)是近十几年来发展起来的一种新的分子标记 ,它是指以少数几个核苷酸 ( 1~ 6个 )为单位多次重复的简单序列 ,以双核苷酸重复最为常见 ,而其中又以 (CA/GT) n 居多。由于微卫星在真核生物基因组中是随机分布的 ,而作为分子遗传标记又有着非常高的多态性和共显性 ,因此在构建遗传连锁图谱时备受青睐 (Brooketal ,1994 )。目前 ,在人类和多种动物中已经构建了以微卫星为主的遗传连锁图谱。但在鲤鱼等水产动物的遗传连锁图谱中 ,微卫星分子标记还较少 (孙效文和梁利群 ,2 0 0 0 )。为了摸索…  相似文献
4.
Strategies for microsatellite isolation: a review   总被引:66,自引:0,他引:66  
In the last few years microsatellites have become one of the most popular molecular markers used with applications in many different fields. High polymorphism and the relative ease of scoring represent the two major features that make microsatellites of large interest for many genetic studies. The major drawback of microsatellites is that they need to be isolated de novo from species that are being examined for the first time. The aim of the present paper is to review the various methods of microsatellite isolation described in the literature with the purpose of providing useful guidelines in making appropriate choices among the large number of currently available options. In addition, we propose a fast and easy protocol which is a combination of different published methods.  相似文献
5.
大熊猫微卫星DNA的筛选及其应用   总被引:62,自引:3,他引:59       下载免费PDF全文
经DpnⅡ限制酶消化大熊猫基因组DNA后,选取150-500bp大小的片段构建了DNA文库。用合成的(CA)(15)探针从这一文库中克隆筛选了10个大熊猫特异的微卫星DNA座位。在测定DNA序列的基础上设计并合成了10个座位特异的引物,以PCR技术扩增了7份抽提自组织细胞和6份抽提自毛发的大熊猫DNA样品。发现除座位g(007)外,其余9个座位均呈多态性,而且所有座位均为偶数碱基的长度变异。对有准确谱系记录的家系的研究结果证明,这10个微卫星DNA座位严格按孟德尔方式遗传。因而这些座位能有效地应用于大熊猫的亲子鉴定,从而为建立各地大熊猫谱系和制定有效的繁殖计划提供了一个有效的遗传学手段。根据这10个微卫星座位的分析,我们澄清了两组未知的父系关系。  相似文献
6.
The utility of RFLP (restriction fragment length polymorphism), RAPD (random-amplified polymorphic DNA), AFLP (amplified fragment length polymorphism) and SSR (simple sequence repeat, microsatellite) markers in soybean germplasm analysis was determined by evaluating information content (expected heterozygosity), number of loci simultaneously analyzed per experiment (multiplex ratio) and effectiveness in assessing relationships between accessions. SSR markers have the highest expected heterozygosity (0.60), while AFLP markers have the highest effective multiplex ratio (19). A single parameter, defined as the marker index, which is the product of expected heterozygosity and multiplex ratio, may be used to evaluate overall utility of a marker system. A comparison of genetic similarity matrices revealed that, if the comparison involved both cultivated (Glycine max) and wild soybean (Glycine soja) accessions, estimates based on RFLPs, AFLPs and SSRs are highly correlated, indicating congruence between these assays. However, correlations of RAPD marker data with those obtained using other marker systems were lower. This is because RAPDs produce higher estimates of interspecific similarities. If the comparisons involvedG. max only, then overall correlations between marker systems are significantly lower. WithinG. max, RAPD and AFLP similarity estimates are more closely correlated than those involving other marker systems.Abbreviations RFLP restriction fragment length plymorphism - RAPD random-amplified polymorphic DNA - AFLP amplified fragment length polymorphism - SSR simple sequence repeat - PCR polymerase chain reaction - TBE Tris-borate-EDTA buffer - MI marker index - SENA sum of effective numbers of alleles  相似文献
7.
新疆8个绵羊品种遗传多样性和系统发生关系的微卫星分析   总被引:60,自引:0,他引:60  
为分析新疆北疆地区主要绵羊品种的遗传多样性和系统发生关系,利用10个微卫星标记,采用PCR扩增,12%非变性聚丙烯酰胺凝胶电泳、Sanguinetti银染法显色,对新疆北疆地区8个品种、1个杂交一代绵羊群体遗传多样性进行了检测,统计了各群体的等位基因组成、平均有效等位基因数(E)和平均基因纯合率,利用等位基因频率计算出各群体的平均遗传杂合度(h)、多态信息含量(PIC)和群体间的遗传距离。利用分子进化遗传分析软件,采用邻结法构建系统发生树;同时根据等位基因频率,利用PHYLIP(3.6)分析软件,采用最大似然法构建系统发生树,应用白举检验估计系统树中结点的白引导值,并进行了系统发生分析。结果表明:10个微卫星位点在9个绵羊群体中的多态信息含量除BMI824、MAF65为低、中度多态外,其余8个微卫星均为高度多态,可作为有效的遗传标记用于各绵羊品种的遗传多样性和系统发生关系的分析;所有绵羊群体的平均PIC(0.5631)、h(0.5721)和E(2.9)均低于国外其他品种的绵羊,其基因多态性和遗传多样性相对贫乏;新疆本地土种阿勒泰羊、哈萨克羊和巴什拜羊与国外引进绵羊品种及混有外血的本地培育品种遗传距离较远,他们聚为不同的两类,各绵羊品种的分子系统发生关系与其来源、育成史、分化及地理分布基本一致。  相似文献
8.
小尾寒羊五个微卫星基因座遗传多态性研究   总被引:60,自引:5,他引:55  
小尾寒羊是我国优良的地方绵羊品种,具有极高的繁殖力,平均每胎产羔2.6只。利用与绵羊高繁殖力主效基因Fec^B和FecX^1连锁的5个微卫星标记(OarAE101,BM1329,BMS2508,TGLA54t TGLA68)对244小尾寒羊母羊进行了遗传检测。用非变性(中性)聚丙烯酰胺凝胶电泳检测同卫星的PCR扩增产物,计算了5个同卫星基因座的等位基因频率,多态信息含量,基因纯合度和杂合度。在小尾寒羊中检测到BM1329有6个等位基因,片段大小为160-180bp,164bp等位基因频率最高(0.6320);检测到OarAE101有9个等位基因,片段大小为97-135bp,97bp等位基因频率最高(0.7930);检测到TGLA54有5个等位基因,片段大小为116-136bp,134bp等位基因频率最高(0.8500);检测到TGLA68有2个等位基因,片段大小为98-100bp,2个等位基因频率相近,检测到BMS2508有6个等位基因,片段大小为93-115bp,99bp等位基因频率最高(0.4795)。BM1329,OarAE101,TGLA54,TGLA68,BMS2508的多态信息含量/基因纯合度/杂合度分别为0.4481/0.4840.0.5160,0.3516/0.6375/0.3625,0.2528/0.7326/0.2674,0.3733/0.5034/0.4966,0.5809/0.3581/0.6419。可见BMS2508的遗传变异最大,TGLA54的遗传变异最小。这些结果可为小尾寒羊种质特性研究提供分子基础数据。  相似文献
9.
微卫星及其应用   总被引:58,自引:7,他引:51       下载免费PDF全文
微卫星是广泛分布于真核生物基因组中的短串联重复序列(1-5bp),具有突变速率快、多态性高等特性,已被广泛应用于生物遗传作图、群体遗传研究、个体间亲缘关系鉴定等方面。简要论述了微卫星的突变、位点的分离、数据的收集、在生物学中的应用及其存在的缺陷。  相似文献
10.
东北虎微卫星DNA遗传标记的筛选及在亲子鉴定中的应用   总被引:56,自引:0,他引:56  
利用18个家猫微卫星基因座,在东北虎(Panthera tigris sibilia)DNA中扩增结果有4个基因座没有产物,8个基因座为单态,6个基因座为多态性。同时利用苏门答腊虎的微卫星序列设计了8对引物,在东北虎DNA中有4对具有多态性。微卫星基因座的多态性百分率为38.5%。在供试的27只东北虎中,发现等位基因间的变异均为偶数碱基长度变化,对有准确谱系记录的个体研究表明,这10个微卫星DNA遗传标记符合孟德尔遗传规律,所以这些微卫星DNA可以有效的应用于东北虎的亲子鉴定。利用这10对多态性引物,我们成功地鉴定了7个父子关系不清的后代。收集的样品包括23只毛发样品和4只血液样品,实验结果表明,毛发和血液样品均可以得到清晰的微卫星条带[动物学报49(1):118—123.2003]。  相似文献
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