Hypervariable 'minisatellite' regions in human DNA

The human genome contains many dispersed tandem-repetitive 'minisatellite' regions detected via a shared 10-15-base pair 'core' sequence similar to the generalized recombination signal (chi) of Escherichia coli. Many minisatellites are highly polymorphic due to allelic variation in repeat copy number in the minisatellite. A probe based on a tandem-repeat of the core sequence can detect many highly variable loci simultaneously and can provide an individual-specific DNA 'fingerprint' of general use in human genetic analysis.     

Forensic application of DNA 'fingerprints'

Many highly polymorphic minisatellite loci can be detected simultaneously in the human genome by hybridization to probes consisting of tandem repeats of the 'core' sequence. The resulting DNA fingerprints produced by Southern blot hybridization are comprised of multiple hypervariable DNA fragments, show somatic and germline stability and are completely specific to an individual. We now show that this technique can be used for forensic purposes; DNA of high relative molecular mass (Mr) can be isolated from 4-yr-old bloodstains and semen stains made on cotton cloth and digested to produce DNA fingerprints suitable for individual identification. Further, sperm nuclei can be separated from vaginal cellular debris, obtained from semen-contaminated vaginal swabs, enabling positive identification of the male donor/suspect. It is envisaged that DNA fingerprinting will revolutionize forensic biology particularly with regard to the identification of rape suspects.     

AFLP技术及其在动物遗传育种中的应用

扩增片段长度多态性(AFLP)被称为“下一代分子标记”。是检测DNA多态性的一种新技术．该技术可靠性强,多态性检出率高,因而被认为是最有效的DNA指纹分析技术,AFLP已广泛应用于分类学、种群遗传学、病理学、DNA指纹分析的研究和建立数量性状基因图谱,成为最主要的遗传标记,论述了AFLP的原理、特点、影响实验成功的关键因素及在动物遗传育种中的应用。     

Estimating genetic diversity and sampling strategy for a wild soybean （Glycine soja） population based on different molecular markers

With the rapid development of the global economy and continued increase in world population, natural environments face serious deterioration and change, which has led to the extinction or severe endangerment of many plant species including important crops…     

草鱼基因组随机扩增多态性引物及多态性位点的筛选

为了建立草鱼基因组DNA多态性分析的指标体系,应用RAPD技术,从180条10碱基随机引物中筛选出了31条能扩增出多态性DNA片段的引物。用这31条多态性引物共扩增出了327条重复性好、带型清晰、分辨率高的谱带。扩增产物的片段大小范围在400—2000bp之间。单一引物扩增条带为5—14条。用这些多态性引物在草鱼基因组DNA中检测到了93个多态性位点,并对这些多态性位点上的等位基因频率进行了统计分析。这31条多态性引物和所检测到的93个多态性位点初步为草鱼基因组的多态性分析提供了可靠的分析指标体系。     

Spontaneous mutation rates to new length alleles at tandem-repetitive hypervariable loci in human DNA

Tandem-repetitive minisatellite regions in vertebrate DNA frequently show substantial allelic variation in the number of repeat units. This variation is thought to arise through processes such as unequal crossover or replication slippage. We show here that the spontaneous mutation rate to new length alleles at extremely variable human minisatellites is sufficiently high to be directly measurable in human pedigrees. The mutation rate at different loci increases with variability in accord with the neutral mutation/random drift hypothesis, and rises to 5% per gamete for the most unstable human minisatellite isolated. Mutations are sporadic, occur with similar frequencies in sperm and oocytes, and can involve the gain or loss of substantial numbers of repeat units, consistent with length changes arising primarily by unequal exchange at meiosis. Germline instability must therefore be taken into account when using hypervariable loci as genetic markers, particularly in pedigree analysis and parenthood testing.     

丁鱼岁遗传多样性的随机扩增多态性DNA分析

利用RAPD分子标记技术对15个丁鱼岁养殖个体的遗传多样性进行分析.选用20个随机引物对其基因组DNA进行PCR扩增,有16个引物可以扩增出稳定且清晰的条带,其中S103、S104和S105为多态引物.16个引物共扩增出65个DNA位点,片段大小在200~3 000 bp之间,其中多态性位点6个,多态位点比例为9.23%.个体间遗传距离在0~0.067 4之间,平均遗传距离为0.028 6.结果显示,丁鱼岁养殖群体的遗传多样性水平较低.     

Positive identification of an immigration test-case using human DNA fingerprints

The human genome contains a set of minisatellites, each of which consists of tandem repeats of a DNA segment containing the 'core' sequence, a putative recombination signal in human DNA. Multiallelic variation in the number of tandem repeats occurs at many of these minisatellite loci. Hybridization probes consisting of tandem repeats of the core sequence detect many hypervariable minisatellites simultaneously in human DNA, to produce a DNA fingerprint that is completely individual-specific and shows somatic and germline stability. These DNA fingerprints are derived from a large number of highly informative dispersed autosomal loci and are suitable for linkage analysis in man, and for individual identification in, for example, forensic science and paternity testing. They can also be used to resolve immigration disputes arising from lack of proof of family relationships. To illustrate the potential for positive or inclusive identification, we now describe the DNA fingerprint analysis of an immigration case, the resolution of which would have been very difficult and laborious using currently available single-locus genetic markers.     

上海地区汉族人单胺氧化酶基因微卫星DNA多态性研究

应用扩增片段长度多态性技术,分析单胺氧化酶基因的ＭＡＯＡ（ＣＡ）。力ＭＡＯＢＩ（ＧＴ）ｎ２个基因座的微卫星ＤＮＡ多态性。在２４６名（男性１２２人,女性１２４人）无血缘关系上海地区汉族人中检出ＭＡＯＡ（ＣＡ）ｎ基因座８－种等位基因（１１０－１２４ｂｐ）和１９种基因型。     

DNA分子技术与鱼类种质资源研究

DNA分子技术为生物种内和种间的比较及遗传多样性提供了一种崭新而又行之有效的途径。简要介绍了Southerm杂交、RFLP、PCR、小卫星和微卫星技术、RAPD、原位PCR和DNA测序等几种DNA分子技术及其在鱼类种质资源研究中的应用情况。概述了我国分子水平上的鱼类种质资源研究现状和进展。     

DNA transformation leads to pilin antigenic variation in Neisseria gonorrhoeae

Many pathogenic bacteria express pili (fimbriae) on their cell surfaces. These structures mediate binding of bacteria to host tissues, and may also be involved in other aspects of pathogenesis. Neisseria gonorrhoeae pili are mainly composed of a single protein, pilin, whose expression is controlled at chromosomal expression loci (pilE). An intact pilin gene and promoter sequences are only found at pilE. Strain MS11 contains two expression sites (pilE1 and pilE2), whereas several of its derivatives and other clinical isolates contain only one. Silent pilin loci (pilS1-pilS7) contain truncated variant pilin genes lacking the promoter and conserved pilin gene sequences. Pilin antigenic variation in N. gonorrhoeae occurs by DNA recombination between one of he silent partial variant gene segments in pilS and an expressed pilin gene in pilE. The recombination reactions are nonreciprocal, and therefore the mechanism has been classified as gene conversion. We report that much of the recombination between pilin loci actually occurs after transformation of living piliated cells by DNA liberated from lysed cells within a population. This constitutes a new molecular mechanism for an antigenic variation system, as well as the first specific function for a DNA transformation system.     

斜带石斑鱼、赤点石斑鱼RAPD和线粒体Cyt b基因序列变异分析

斜带石斑鱼和赤点石斑鱼为具有重要经济价值的海水鱼类。对这两种石斑鱼进行了随机扩增多态DNA(RAPD)和线粒体细胞色素b(Cyt b)基因序列变异分析。计算多态位点百分率、基因多样性和香农信息指数等遗传参数,以此评估种内遗传变异水平;通过统计变异位点、平均核苷酸差异数、核苷酸多样性以及种间平均每位点核苷酸替代数进行基因序列变异分析,并构建UPGMA系统树。结果表明:斜带石斑鱼的遗传多样性水平高于赤点石斑鱼,这可能与它们在种内特定遗传结构、分布范围大小、自然资源状况的差异有关;斜带石斑鱼和赤点石斑鱼之间检测到的种特异RAPD条带以及基因序列的变异,可作为种间分子鉴定标记。     

Genetic evidence against panmixia in the European eel

The panmixia hypothesis--that all European eel (Anguilla anguilla) migrate to the Sargasso Sea for reproduction and comprise a single, randomly mating population--is widely accepted. If true, then this peculiar life history strategy would directly impact the population genetics of this species, and eels from European and north African rivers should belong to the same breeding population through the random dispersal of larvae. To date, the panmixia hypothesis has remained unchallenged: genetic studies realized on eel's mitochondrial DNA failed to detect any genetic structure; and a similar lack of structure was found using allozymes, with the exception of clinal variation imposed by selection. Here we have used highly polymorphic genetic markers that provide better resolution to investigate genetic structure in European eel. Analysis of seven microsatellite loci among 13 samples from the north Atlantic, the Baltic Sea and the Mediterranean Sea basins reveals that there is global genetic differentiation. Moreover, pairwise Cavalli-Sforza and Edwards chord distances correlate significantly with coastal geographical distance. This pattern of genetic structure implies non-random mating and restricted gene flow among eels from different sampled locations, which therefore refute the hypothesis of panmixia. Consequently, the reproductive biology of European eel must be reconsidered.     

野生和人工选育黄河鲤遗传多样性的ISSR分析

利用ISSR分子标记技术,对采集于长垣天然文岩渠(野生群体)、河南省水产科学研究院(黄河品系Ⅰ群体)、小浪底水库(野生群体)、河南黄河鲤鱼良种场(豫选黄河鲤群体)的4个群体黄河鲤的120个个体的遗传多样性进行了分析.8个ISSR引物共获得64个扩增位点,其中多态位点31个,多态位点比例为48.44%.4个群体的多态位点比例分别为14.06%,32.81%,34.38%和20.31%,遗传距离分别为0.0576、0.1657、0.1674和0.0800,Nei基因多样性分别为0.0538、0.1099、0.1308和0.0723,Shannon信息指数分别为0.0784、0.1669、0.1923和0.1072.文岩渠群体与黄河品系Ⅰ群体遗传距离最远(0.1717),与小浪底水库群体遗传距离最近(0.0558).表明这些群体间发生了较弱的遗传分化.UPGMA聚类结果为文岩渠群体与小浪底群体先聚在一起,其次是与豫选黄河鲤,最后与黄河鲤品系Ⅰ相聚.用AMOVA进行遗传变异方差分析得到遗传变异固定指数(Φst=0.0179,P0.05),显示黄河鲤大部分变异(98.21%)发生在群体内,群体间变异较小(1.79%).说明人工选育群体的遗传结构尚未发生明显变化.     

林木分子育种研究的基因组学信息资源述评

分子育种是指利用与性状相关的DNA标记进行选育,也称标记辅助选择或标记辅助育种,广义上还包括基因工程育种和基因组学辅助育种。林木分子育种为早期选择和加速育种提供了极具潜力的高效手段。笔者对林木分子育种研究的基因组学信息资源进行了进展综述和前景展望。近30年来,林木分子标记技术从早期的低通量方法发展到目前基于微阵列芯片和新一代测序的高通量技术,如测序分型、转录组测序、重测序、扩增子测序和外显子组测序等,并广泛用于连锁作图、关联分析和基因组选择等林木性状相关的DNA变异检测研究。随着2006年毛果杨基因组序列的发表,已有50余个树种完成了基因组测序。基于连锁作图和关联研究检测了林木10余个属生长、材性和抗逆及非木质产品品质等性状相关的大量基因组位点,主要趋势表现为：① 表型广泛,涵盖经济性状、生理指标和代谢成分等;②标记数量成千上万甚至上百万,覆盖全基因组;③转录组和降解组等多组学的分子变异开始应用;④ 利用大群体以提高位点检测的精度;⑤ 重视环境的影响,大田试验设置多个地点,解析QTL与环境、年份的互作效应;⑥ 结合参考基因组序列和/或转录组差异表达基因进一步挖掘性状相关的候选基因,建立了桉属、松属和云杉属等主要造林树种的基因组选择模型。此外,积累了泛基因组、相关软件和算法、功能基因、基因组编辑技术及网站和数据库等其他信息资源。林木分子育种面临的挑战主要包括：① 如何获得稳定性好的性状相关基因组位点和基因组选择（GS）模型;② 缺乏自动化、无损和高通量的表型测定技术;③对大基因组的针叶树和一些多倍体树种,仍难获得高质量的基因组序列;④ 标记辅助选择增加了常规育种之外的费用,且存在不确定性;⑤多数树种的加速育种仍较困难。后基因组时代的林木分子育种将有效结合到常规育种程序中,显著促进遗传增益的提高。     

东海野生灰鲳mtDNA D-loop序列遗传变异分析

以东海区野生灰鲳背部肌肉的线粒体DNA为模板,采用PCR技术对7个个体的D-loop序列进行了扩增,通过对PCR产物进行双向测序,最终得到了471 bp的核苷酸片断(除去两端部分序列)。用DNAMAN6.0软件进行了排序比较发现,东海野生灰鲳的D-loop序列同源性高达99.48%;用DNASP4.0软件分析得知,7个序列共有6个单倍型(在GenBank登录号:GU970085-GU970087,GU970089-GU970091),在7个个体中,共检测到14个变异位点,包括13个转换和1个颠换位点。运用MEGA4.0软件计算出了不同个体间的遗传距离,并据此构建了MP和NJ系统树。用DNASP4.0软件计算出的多态位点数为14,核苷酸多样性指数和平均核苷酸差异数分别为0.009 71和4.571。研究结果表明,东海野生灰鲳的D-loop序列个体变异程度并不丰富。     

分子标记在杨树遗传改良中的应用

介绍了杨树遗传改良中常用的三种分子标记;ＲＦＬＰ、ＲＡＰＤ和ＡＦＬＰ,并回顾了分子标记在杨树遗传改良中的应用;构建无性系指纹图谱,构建遗传图谱,群体遗传变异和系统进化及分类,杂种鉴定,数量性状基因定位以及分子标记辅助选择育种等。     

Variations in DNA elucidate molecular networks that cause disease

Identifying variations in DNA that increase susceptibility to disease is one of the primary aims of genetic studies using a forward genetics approach. However, identification of disease-susceptibility genes by means of such studies provides limited functional information on how genes lead to disease. In fact, in most cases there is an absence of functional information altogether, preventing a definitive identification of the susceptibility gene or genes. Here we develop an alternative to the classic forward genetics approach for dissecting complex disease traits where, instead of identifying susceptibility genes directly affected by variations in DNA, we identify gene networks that are perturbed by susceptibility loci and that in turn lead to disease. Application of this method to liver and adipose gene expression data generated from a segregating mouse population results in the identification of a macrophage-enriched network supported as having a causal relationship with disease traits associated with metabolic syndrome. Three genes in this network, lipoprotein lipase (Lpl), lactamase beta (Lactb) and protein phosphatase 1-like (Ppm1l), are validated as previously unknown obesity genes, strengthening the association between this network and metabolic disease traits. Our analysis provides direct experimental support that complex traits such as obesity are emergent properties of molecular networks that are modulated by complex genetic loci and environmental factors.     

Construction of a genetic linkage map for cotton based on SRAP

DNA markers have been widely used in construction of molecular genetic linkage maps in plants. The first genetic linkage map of cotton was constructed by Reinish in 1994 using RFLP (restriction fragment length polymorphism)[1], which included 705 polymorphic loci on 41 linkage groups with a total length of 4675 cM. Afterwards, several genetic linkage maps were constructed[2—7], but no map is comparable to this one in marker density. A high-density genetic linkage map could be applied effec…     

A preliminary study on S．asotus linnaeus and S．soldatovi meridionalis Chen by DNA fingerprinting

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