单核苷酸多态性及其在鸡QTL定位上的应用

单核苷酸多态性是指DNA序列上的单个碱基变异,它具有分布广、多态信息含量大、易于检测和统计分析等优点,能较好用于基因图谱构建和数量性状QTL定位研究,被称为继RFLP和微卫星标记之后的第3代基因遗传标记。本文综述了单核苷酸多态性的性质及检测技术、利用候选基因SNP进行鸡QTL定位研究的现状,并对未来SNP的应用前景进行了展望。Abstract:Single nucleotide polymorphism (SNP) refers to the change of single nucleotide in DNA sequence.Because of its high density in genomes and easy in detection and analysis statistically,SNP can be used in genetic linkage map construction and QTL mapping.Here,the characters and detecting technology of SNP,as well as the status and foreground of the use of candidate gene SNP in chicken QTL mapping are introduced.     

A multiplexer liquidchip technology for detecting single nucleotide polymorphisms from metabolism of anti-thrombotic drugs in dried blood spots on filter paper

A single nucleotide polymorphism （SNP） is the most fre quent type of variation in the genome. There are around 10 million SNPs that have been identified in the human genome [1]. Because SNPs are highly conserved throughout evolu tion and within a population, the map of SNPs serves as an excellent genotypic marker for research. The elucidation of SNP information will contribute to an individual＇s suscepti bility to disease and responsiveness to drug toxicity and medical intervention [2,3]. Nowadays, a variety of techni ques have been used to perform SNP genotyping, but these techniques required whole blood as the sample. Dried blood spot （DBS） specimens require less material and are substan tially more stable （several months at room temperature） than whole blood [4]. Thus, the simplicity of sample preparation, long time storage and convenient transport make DBS to be a costeffective and suitable alternative tool for collecting blood sample.     

Identification of Risk Pathways and Functional Modules for Coronary Artery Disease Based on Genome-wide SNP Data

Coronary artery disease(CAD) is a complex human disease, involving multiple genes and their nonlinear interactions, which often act in a modular fashion. Genome-wide single nucleotide polymorphism(SNP) profiling provides an effective technique to unravel these underlying genetic interplays or their functional involvements for CAD. This study aimed to identify the susceptible pathways and modules for CAD based on SNP omics. First, the Wellcome Trust Case Control Consortium(WTCCC) SNP datasets of CAD and control samples were used to assess the jointeffect of multiple genetic variants at the pathway level, using logistic kernel machine regression model. Then, an expanded genetic network was constructed by integrating statistical gene–gene interactions involved in these susceptible pathways with their protein–protein interaction(PPI)knowledge. Finally, risk functional modules were identified by decomposition of the network. Of 276 KEGG pathways analyzed, 6 pathways were found to have a significant effect on CAD. Other than glycerolipid metabolism, glycosaminoglycan biosynthesis, and cardiac muscle contraction pathways, three pathways related to other diseases were also revealed, including Alzheimer's disease, non-alcoholic fatty liver disease, and Huntington's disease. A genetic epistatic network of 95 genes was further constructed using the abovementioned integrative approach. Of 10 functional modules derived from the network, 6 have been annotated to phospholipase C activity and cell adhesion molecule binding, which also have known functional involvement in Alzheimer's disease.These findings indicate an overlap of the underlying molecular mechanisms between CAD and Alzheimer's disease, thus providing new insights into the molecular basis for CAD and its molecular relationships with other diseases.     

Progress in the Study of Molecular Genetic Improvements of Poplar in China

The poplar is one of the most economically important and intensively studied tree species owing to its wide application in the timber industry and as a model material for the study of woody plants. The natural resource of poplars in China is replete. Over the past 10 years, the application of molecular biological techniques to genetic improvements in poplar species has been widely studied in China. Recent advances in molecular genetic improvements of poplar, including cDNA library construction, gene cloning and identification, genetic engineering, gene expression, genetic linkage map construction, mapping of quantitative trait loci （QTL） and molecular-assisted selection, are reviewed in the present paper. In addition, the application of modern biotechnology to molecular improvements in the genetic traits of the poplar and some unsolved problems are discussed.     

小麦SSR标记的发展及应用

微卫星是以1～6个碱基为基本单元的串联重复序列,由于具有共显性、多态性高和容易用PCR方法检测等特点,是非常有用的遗传标记。在小麦中,SSR标记已广泛应用于遗传图谱的构建、遗传多样性、品种及基因型鉴定、目的基因,以及QTL的标记和标记辅助选择育种。 Abstract:Microsatellites are simple,tandemly repeated one to six nucleotide sequence motifs.They are very useful as genetic markers because they are co-dominant,detect high levels of allelic diversity,and are easily assayed by the polymerase chain reaction ( PCR ).In wheat,SSR markers have been applied to genetic mapping,detection of genetic diversity,identification of varieties and genotypes,gene tagging,QTL analysis,and marker-assisted selection.     

QTL mapping of economically important traits in Silkworm (Bombyx mori)

The construction of linkage map is both a funda-mental research area and an important aspect of gene analyses in genetics. It provides the guidelines for breeding. A sound linkage map is also necessary for further genetic analysis. In recent years, great and rapid progress has been made in molecular biology, which enables fingerprinting of organisms at the ge-nomic level. Many molecular marker techniques have been well established. Heartening progress has been made in many organisms in the co…     

林木遗传连锁图谱构建研究进展与发展方向

本文就目前国内外林木连锁遗传图谱领域的研究进展进行了综述,指出了该领域研究中存在的主要问题,即一方面是作图个体的数量有限,另一方面是采用的标记以随机标记为主,导致了建成的图谱以及利用图谱获得的数量性状基因位点（QTLs）信息具有杂交组合特异性,造成了QTLs的可信度和在林木遗传改良以及标记辅助选择中的实用性降低等现象。针对存在的问题,讨论了根据林木生物学特点选择合适遗传标记的意义,指出进行林木比较作图研究的重要性和必要性。文中接着较为详尽地介绍了国外重要林木表达序列标签（EST）测序项目的研究进展,论述了功能已知和种间高度保守的表达序列标签多态性（ESTP）标记的由来,阐述了获得ESTP标记的主要方法,并指出应当利用ESTP标记进行林木遗传图谱构建、QTL定位和比较作图的研究。文中最后讨论了未来林木遗传图谱构建和QTL定位研究的发展方向,并探讨了我国在该领域取得重大进展的突破口,指出我国应首先进行杨树尤其是中国乡土杨树树种该方面的研究。 Abstract:The research progress in genetic linkage map construction of forest tree species both at home and abroad were reviewed in the paper.Two main problems involved in the field were discussed.One was the limitation of the number of individuals of mapping populations and the other was the random markers mostly employed by the majority of studies.These problems have resulted in crossing combination specificity in the constructed maps and the QTLs located on the basis of the maps.As a result,the QTLs discovered up to now have low credibility and poor practicability in marker-assisted selection.Therefore considering the biological characteristics of forest tree species,the selection of the most suitable genetic markers is crucial to obtain a high quality genetic linkage map,and it is both important and necessary to carry out comparative genetic mapping.Progress in the ongoing expressed sequence tag (EST) sequencing projects were summarized and EST polymorphism (ESTP),the most informative and highly conservative marker with known function,as well as the main ESTP detection techniques were elaborated.It was pointed out that ESTP markers should be integrated into the present studies of genetic linkage map construction,QTL mapping and genome comparative mapping.Finally the future prospects in the fields of genetic linkage map and QTL mapping were discussed.In China,Such studies around Populus,especially in the local Populus species should make a breakthrough in the related fields.     

单核苷酸多态性检测方法的新进展

单核苷酸多态性（single nucleotide polymorphism,SNP）是第三代遗传标记,在基因定位、遗传疾病和人类起源等理论研究中具有重大意义, 在抗药性或药物过敏反应中扮演着极其重要的角色,正逐步成为分子诊断、临床检验、新药研发的重要手段。随着人类基因组测序的完成,SNP分型和发现成为遗传和生命医学领域研究的热点之一。近年来,SNP的检测方法层出不穷,发展很快。文章综述和分析了几种新建立的SNP检测方法,包括基因芯片、分子探针、荧光偏振、荧光共振、质谱和磁性颗粒分析。在生物化学、工程学和分析软件等方面取得突破的基础上,有望建立灵敏准确、简便易行、高通量、低费用的SNP技术。Abstract: Single nucleotide polymorphism (SNP) is the third generation genetic marker. SNP detection now is becoming increasingly important means in molecular diagnostics, clinical assay and novel drug development. It plays an essential role in drug resistance and anaphylactic reaction and has the importamce in theoretical studies of gene location, hereditary diseases and human origin. With the accomplishment of human genome sequencing, the genotyping and discovering of SNP are becoming hot subjects in genetics and biomedicine researches. The methods for SNP detection were renewed rapidly and developed fast in past few years. In this review, several newly established detection methods including gene chip, molecular probe, fluorescence polarization and resonance, mass spectrometry, and bacterial magnetic particle are discussed. It could be expected that an accurate and sensitive, simple and easy-to-handle SNP technology with low cost and high throughput will be available on the basis of research breakthroughs of biochemistry, engineering and analytic software.     

单核苷酸多态性在医学领域的应用

作为第三代DNA遗传标记,单核苷酸多态性(SNP)标记已经成为基因功能研究、寻找疾病基因和基因型鉴定、药物基因组学的主要研究手段,在高危群体的发现、疾病相关基因的鉴定、药物的设计和测试以及分子生物学的基础研究等方面发挥了重要作用。近年来SNP在法医鉴定等领域也有广泛应用。     

SNP marker diversity in common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.)

Single nucleotide polymorphism (SNP) markers have become a genetic technology of choice because of their automation and high precision of allele calls. In this study, our goal was to develop 94 SNPs and test them across well-chosen common bean (Phaseolus vulgaris L.) germplasm. We validated and accessed SNP diversity at 84 gene-based and 10 non-genic loci using KASPar technology in a panel of 70 genotypes that have been used as parents of mapping populations and have been previously evaluated for SSRs. SNPs exhibited high levels of genetic diversity, an excess of middle frequency polymorphism, and a within-genepool mismatch distribution as expected for populations affected by sudden demographic expansions after domestication bottlenecks. This set of markers was useful for distinguishing Andean and Mesoamerican genotypes but less useful for distinguishing within each gene pool. In summary, slightly greater polymorphism and race structure was found within the Andean gene pool than within the Mesoamerican gene pool but polymorphism rate between genotypes was consistent with genepool and race identity. Our survey results represent a baseline for the choice of SNP markers for future applications because gene-associated SNPs could themselves be causative SNPs for traits. Finally, we discuss that the ideal genetic marker combination with which to carry out diversity, mapping and association studies in common bean should consider a mix of both SNP and SSR markers.     

Single nucleotide polymorphism discovery in barley using autoSNPdb

Molecular markers are used to provide the link between genotype and phenotype, for the production of molecular genetic maps and to assess genetic diversity within and between related species. Single nucleotide polymorphisms (SNPs) are the most abundant molecular genetic marker. SNPs can be identified in silico , but care must be taken to ensure that the identified SNPs reflect true genetic variation and are not a result of errors associated with DNA sequencing. The SNP detection method autoSNP has been developed to identify SNPs from sequence data for any species. Confidence in the predicted SNPs is based on sequence redundancy, and haplotype co-segregation scores are calculated for a further independent measure of confidence. We have extended the autoSNP method to produce autoSNPdb, which integrates SNP and gene annotation information with a graphical viewer. We have applied this software to public barley expressed sequences, and the resulting database is available over the Internet. SNPs can be viewed and searched by sequence, functional annotation or predicted synteny with a reference genome, in this case rice. The correlation between SNPs and barley cultivar, expressed tissue type and development stage has been collated for ease of exploration. An average of one SNP per 240 bp was identified, with SNPs more prevalent in the 5' regions and simple sequence repeat (SSR) flanking sequences. Overall, autoSNPdb can provide a wealth of genetic polymorphism information for any species for which sequence data are available.     

Development of single nucleotide polymorphism (SNP) markers for use in commercial maize (Zea mays L.) germplasm

The development of single nucleotide polymorphism (SNP) markers in maize offers the opportunity to utilize DNA markers in many new areas of population genetics, gene discovery, plant breeding and germplasm identification. However, the steps from sequencing and SNP discovery to SNP marker design and validation are lengthy and expensive. Access to a set of validated SNP markers is a significant advantage to maize researchers who wish to apply SNPs in scientific inquiry. We mined 1,088 loci sequenced across 60 public inbreds that have been used in maize breeding in North America and Europe. We then selected 640 SNPs using generalized marker design criteria that enable utilization with several SNP chemistries. While SNPs were found on average every 43 bases in 1,088 maize gene sequences, SNPs that were amenable to marker design were found on average every 623 bases; representing only 7% of the total SNPs discovered. We also describe the development of a 768 marker multiplex assay for use on the Illumina^® BeadArray? platform. SNP markers were mapped on the IBM2 intermated B73 × Mo17 high resolution genetic map using either the IBM2 segregating population, or segregation in multiple parent-progeny triplets. A high degree of colinearity was found with the genetic nested association map. For each SNP presented we give information on map location, polymorphism rates in different heterotic groups and performance on the Illumina^® platform.     

Genomic growth curves of an outbred pig population

In the current post-genomic era, the genetic basis of pig growth can be understood by assessing SNP marker effects and genomic breeding values (GEBV) based on estimates of these growth curve parameters as phenotypes. Although various statistical methods, such as random regression (RR-BLUP) and Bayesian LASSO (BL), have been applied to genomic selection (GS), none of these has yet been used in a growth curve approach. In this work, we compared the accuracies of RR-BLUP and BL using empirical weight-age data from an outbred F2 (Brazilian Piau X commercial) population. The phenotypes were determined by parameter estimates using a nonlinear logistic regression model and the halothane gene was considered as a marker for evaluating the assumptions of the GS methods in relation to the genetic variation explained by each locus. BL yielded more accurate values for all of the phenotypes evaluated and was used to estimate SNP effects and GEBV vectors. The latter allowed the construction of genomic growth curves, which showed substantial genetic discrimination among animals in the final growth phase. The SNP effect estimates allowed identification of the most relevant markers for each phenotype, the positions of which were coincident with reported QTL regions for growth traits.     

Discovery and genetic mapping of single nucleotide polymorphisms in candidate genes for pathogen defence response in perennial ryegrass (<Emphasis Type="Italic">Lolium perenne</Emphasis> L.)

Susceptibility to foliar pathogens commonly causes significant reductions in productivity of the important temperate forage perennial ryegrass. Breeding for durable disease resistance involves not only the deployment of major genes but also the additive effects of minor genes. An approach based on in vitro single nucleotide polymorphism (SNP) discovery in candidate defence response (DR) genes has been used to develop potential diagnostic genetic markers. SNPs were predicted, validated and mapped for representatives of the pathogenesis-related (PR) protein-encoding and reactive oxygen species (ROS)-generating gene classes. The F(1)(NA(6) x AU(6)) two-way pseudo-test cross population was used for SNP genetic mapping and detection of quantitative trait loci (QTLs) in response to a crown rust field infection. Novel resistance QTLs were coincident with mapped DR gene SNPs. QTLs on LG3 and LG7 also coincided with both herbage quality QTLs and candidate genes for lignin biosynthesis. Multiple DR gene SNP loci additionally co-located with QTLs for grey leaf spot, bacterial wilt and crown rust resistance from other published studies. Further functional validation of DR gene SNP loci using methods such as fine-mapping and association genetics will improve the efficiency of parental selection based on superior allele content.     

尼罗罗非鱼ghrelin基因的多态性及其与生长性状相关SNP位点的筛选

为了研究尼罗罗非鱼(Oreochromis niloticus)生长激素促分泌素基因(ghrelin)的多态性及其与生长的相关性, 研究以两个尼罗罗非鱼群体(快长群体和基础群体)的DNA样本各40份为模板, 通过PCR扩增和测序获得ghrelin基因序列。通过Dnasp v5和MEGA 5.0分析序列多态性、筛选有效SNP 位点; 采用Snapshot法对两个群体子代ghrelin基因中SNP位点进行基因分型, 然后分析SNP位点基因型与生长性状的相关性。结果表明, 快长群体ghrelin基因中的单核苷酸变异位点数(S)比基础群体要少, 而核苷酸多态性(Pi)和平均核苷酸差异数(K)要略高于基础群体。共筛得3个有效SNP 位点(S1、S2和S3), 均分布于第1个内含子中。遗传结构分析表明, 3个SNP 位点在两个群体的子代中均为低度多态性位点(PIC0.25), 但处于Hardy-Weinberg平衡(P0.05);快长群体子代中3个SNP 位点的观测杂合度、期望杂合度和多态信息含量等遗传多样性参数均小于基础群体子代的相应值, 3个SNP 位点的遗传多样性参数、基因型和基因频率在同一群体中高度一致, SNP 位点之间完全连锁。两个群体子代中3个SNP 位点处的优势基因型相同, 但快长群体子代中优势基因型频率要明显大于基础群体子代中相应基因型频率。对两个群体子代的生长性状与SNP基因型进行关联性分析的结果表明,尼罗罗非鱼个体的多项生长指标(体重、体长、体高、头长和尾柄高等)在不同基因型中存在显著差异(S1:GG AG, S2:TT AT, S3:AA AT)(P0.05)。D1双倍型(S1:GG, S2:TT, S3:AA)所对应的尼罗罗非鱼个体的多项生长指标(体重、体长、体高、头长和尾柄高等)显著高于D2双倍型(S1:AG, S2:AT, S3:AT)。以上结果表明, 尼罗罗非鱼ghrelin基因3个SNP 位点完全连锁, D1双倍型与快长性状密切相关, 可作为尼罗罗非鱼分子标记辅助育种的候选标记。     

SNP Haplotype Mapping in a Small ALS Family

The identification of genes for monogenic disorders has proven to be highly effective for understanding disease mechanisms, pathways and gene function in humans. Nevertheless, while thousands of Mendelian disorders have not yet been mapped there has been a trend away from studying single-gene disorders. In part, this is due to the fact that many of the remaining single-gene families are not large enough to map the disease locus to a single site in the genome. New tools and approaches are needed to allow researchers to effectively tap into this genetic gold-mine. Towards this goal, we have used haploid cell lines to experimentally validate the use of high-density single nucleotide polymorphism (SNP) arrays to define genome-wide haplotypes and candidate regions, using a small amyotrophic lateral sclerosis (ALS) family as a prototype. Specifically, we used haploid-cell lines to determine if high-density SNP arrays accurately predict haplotypes across entire chromosomes and show that haplotype information significantly enhances the genetic information in small families. Panels of haploid-cell lines were generated and a 5 centimorgan (cM) short tandem repeat polymorphism (STRP) genome scan was performed. Experimentally derived haplotypes for entire chromosomes were used to directly identify regions of the genome identical-by-descent in 5 affected individuals. Comparisons between experimentally determined and in silico haplotypes predicted from SNP arrays demonstrate that SNP analysis of diploid DNA accurately predicted chromosomal haplotypes. These methods precisely identified 12 candidate intervals, which are shared by all 5 affected individuals. Our study illustrates how genetic information can be maximized using readily available tools as a first step in mapping single-gene disorders in small families.     

A rapid, microplate SNP genotype assay for the leptinob allele

A nonsense mutation in the mouse leptin gene causes genetic obesity. As a result of extensive research in the field of obesity, the use of leptinob mice is widespread. This mutation renders mice sterile, creating the need to breed heterozygous mice. For this reason, leptinob genotyping is necessary. To date, gel-based assays have been used for genotyping. Using the Invader Plus assay for single nucleotide polymorphism (SNP) detection, we have developed a gel-free microplate SNP assay for genotyping leptinwt and leptinob alleles.