用商品群作为参考系构建猪的微卫星连锁图谱

由 19头杂种公猪 [皮特兰× (皮特兰×汉普夏 ) ]、5 2头杂种母猪 [Leicoma× (大约克×长白 ) ]及其 332头后代组成的商品群作为参考系 ,选择 172个微卫星标记和 3个Ⅰ类标记 (RYR1、PIT1、PRKAG3)对参考系的个体进行遗传标记分型 ,应用CRIMAP(2 4 )构建猪的整个基因组微卫星连锁图谱。采用多重PCR方法对微卫星标记进行扩增 ,用ABI 377测序仪进行电泳分离 ,应用Genescan 3 0和Genotyper 2 0软件进行基因型检测。 3个Ⅰ类标记用PCR RFLP技术进行分型。CRIMAP程序分析表明 :所构建的猪常染色体性平均连锁图谱的总长度为 2 36 8 7cM ,X染色体的长度为 14 3 10cM ,遗传标记的平均间距为 16 3cM ,亲本的微卫星标记座位的杂合度平均为 0 70。此连锁图谱的构建将为商品猪群的生长、胴体、肉质以及繁殖性状的QTL扫描打下基础。     

Development of Polymorphic EST Markers Suitable for Genetic Linkage Mapping of Catfish

Expressed sequence tag (EST) markers are important for gene mapping and for marker-assisted selection (MAS). To develop EST markers for use in catfish gene mapping, 100 randomly picked complementary DNAs from the channel catfish (Ictalurus punctatus) pituitary library were sequenced. The EST sequences were used to design primers to amplify channel catfish and blue catfish (I. furcatus) genomic DNAs. Polymerase chain reaction products of the ESTs were analyzed to determine length polymorphism between the channel catfish and blue catfish. Eleven polymorphic EST markers were identified. Five of the 11 EST markers were from known genes and the other six were from unidentified ESTs. Seven ESTs were found to be associated with microsatellite sequences. Analysis of channel catfish gene sequences indicated highly biased codon usage, with 16 codons being preferably used. These codons were more preferably used in highly expressed ribosomal protein genes and in highly expressed pituitary hormone genes. G/C-rich codons are less used in channel catfish than those in other vertebrates suggesting AT-richness of the channel catfish genome. Received June 29, 1998; accepted March 29, 1999.     

Integration of Retrotransposons-Based Markers in a Linkage Map of Barley

A deeper understanding of random markers is important if they are to be employed for a range of objectives. The sequence specific amplified polymorphism (S-SAP) technique is a powerful genetic analysis tool which exploits the high copy number of retrotransposon long terminal repeats (LTRs) in the plant genome. The distribution and inheritance of S-SAP bands in the barley genome was studied using the Steptoe × Morex (S × M) double haploid (DH) population. Six S-SAP primer combinations generated 98 polymorphic bands, and map positions were assigned to all but one band. Eight putative co-dominant loci were detected, representing 16 of the mapped markers. Thus at least 81 of the mapped S-SAP loci were dominant. The markers were distributed along all of the seven chromosomes and a tendency to cluster was observed. The distribution of S-SAP markers over the barley genome concurred with the knowledge of the high copy number of retrotransposons in plants. This experiment has demonstrated the potential for the S-SAP technique to be applied in a range of analyses such as genetic fingerprinting, marker assisted breeding, biodiversity assessment and phylogenetic analyses.     

Physical Mapping of 49 Microsatellite Markers on Chromosome 19 and Correlation with the Genetic Linkage Map

We have regionally localized 49 microsatellite markers developed by Généthon using a panel of previously characterized somatic cell hybrids that retain fragments from chromosome 19. The tight correlation observed between the physical and the genetic orders of the microsatellites provide cytogenetic anchorages to the genetic map data. We propose a position for the centromere just above D19S415, from the study of two hybrids, each of which retains one of the two derivatives of a balanced translocation t(1;19)(q11;q11). Microsatellites, which can be identified by a standard PCR protocol, are useful tools for the localization of disease genes and for the establishment of YAC or cosmid contigs. These markers can also judiciously be used for the characterization of new hybrid cell line panels. We report such a characterization of 11 clones, 8 of which were obtained by irradiation-fusion. Using the whole hybrid panel, we were able to define the order of 12 pairs of genetically colocalized microsatellites. As examples of gene mapping by the combined use of microsatellites and hybrid cell lines, we regionally assigned the PVS locus between the 19q13.2 markers D19S417 and D19S423 and confirmed the locations of fucosyltransferase loci FUT1, FUT2, and FUT5.     

基于转录组测序的大熊猫多态性微卫星标记筛选

结合已公布的大熊猫Ailuropoda melanoleuca基因组和本实验室所测6只大熊猫的转录组数据,筛选多态性微卫星位点并分析其组成及特征。结果显示:共获得326个多态性微卫星位点,其中二碱基多态性微卫星最多,共228个,占69.93%;三、四、五、六碱基所占比例分别为9.51%、14.11%、5.21%、1.22%。根据分析结果中缺失率与标准差2项指标以及位点序列长度,选取20个多态性二碱基微卫星位点,用于25只大熊猫个体血液DNA进行PCR验证并做后续分析。结果表明:不同位点的等位基因数为2～8,平均等位基因数为3.70,观测杂合度、期望杂合度分别为0～1.000、0.280～0.784,平均值分别为0.472和0.532。在Bonferroni校正后,证实4个位点显著偏离哈迪-温伯格平衡,所有位点未观察到显著连锁不平衡(P>0.01)。20个位点多态信息含量(PIC)在0.246～0.734,其中具有高度多态性的位点9个(PIC>0.50),11个位点呈中度多态性(0.25相似文献   

High-Throughput SNP Discovery and Genotyping for Constructing a Saturated Linkage Map of Chickpea (Cicer arietinum L.)

The present study reports the large-scale discovery of genome-wide single-nucleotide polymorphisms (SNPs) in chickpea, identified mainly through the next generation sequencing of two genotypes, i.e. Cicer arietinum ICC4958 and its wild progenitor C. reticulatum PI489777, parents of an inter-specific reference mapping population of chickpea. Development and validation of a high-throughput SNP genotyping assay based on Illumina''s GoldenGate Genotyping Technology and its application in building a high-resolution genetic linkage map of chickpea is described for the first time. In this study, 1022 SNPs were identified, of which 768 high-confidence SNPs were selected for designing the custom Oligo Pool All (CpOPA-I) for genotyping. Of these, 697 SNPs could be successfully used for genotyping, demonstrating a high success rate of 90.75%. Genotyping data of the 697 SNPs were compiled along with those of 368 co-dominant markers mapped in an earlier study, and a saturated genetic linkage map of chickpea was constructed. One thousand and sixty-three markers were mapped onto eight linkage groups spanning 1808.7 cM (centiMorgans) with an average inter-marker distance of 1.70 cM, thereby representing one of the most advanced maps of chickpea. The map was used for the synteny analysis of chickpea, which revealed a higher degree of synteny with the phylogenetically close Medicago than with soybean. The first set of validated SNPs and map resources developed in this study will not only facilitate QTL mapping, genome-wide association analysis and comparative mapping in legumes but also help anchor scaffolds arising out of the whole-genome sequencing of chickpea.     

Construction of a High-Density Microsatellite Genetic Linkage Map and Mapping of Sexual and Growth-Related Traits in Half-Smooth Tongue Sole (Cynoglossus semilaevis)

High-density genetic linkage maps of half-smooth tongue sole were developed with 1007 microsatellite markers, two SCAR markers and an F1 family containing 94. The female map was composed of 828 markers in 21 linkage groups, covering a total of 1447.3 cM, with an average interval 1.83 cM between markers. The male map consisted of 794 markers in 21 linkage groups, spanning 1497.5 cM, with an average interval of 1.96 cM. The female and male maps had 812 and 785 unique positions, respectively. The genome length of half-smooth tongue sole was estimated to be 1527.7 cM for the females and 1582.1 cM for the males. Based on estimations of the map lengths, the female and male maps covered 94.74 and 94.65% of the genome, respectively. The consensus map was composed of 1007 microsatellite markers and two SCAR markers in 21 linkage groups, covering a total of 1624 cM with an average interval of 1.67 cM. Furthermore, 159 sex-linked SSR markers were identified. Five sex-linked microsatellite markers were confirmed in their association with sex in a large number of individuals selected from different families. These sex-linked markers were mapped on the female map LG1f with zero recombination. Two QTLs that were identified for body weight, designated as We-1 and We-2, accounted for 26.39% and 10.60% of the phenotypic variation. Two QTLs for body width, designated Wi-1 and Wi-2, were mapped in LG4f and accounted for 14.33% and 12.83% of the phenotypic variation, respectively. Seven sex-related loci were mapped in LG1f, LG14f and LG1m by CIM, accounting for 12.5–25.2% of the trait variation. The results should prove to be very useful for improving growth traits using molecular MAS.     

微卫星标记对资源猪群的遗传分析和连锁图谱构建

在猪数量性状位点的定位研究中,标记的使用和图谱的构建是很重要的。本研究从猪的第4、6、7、8和13染色体上选取39个微卫星标记,在来源于约克夏和梅山214头猪组成的资源群中,分析了遗传特征并构建了图谱。研究表明,平均等位基因数、平均观察杂合度(Ho)和平均多态信息含量(PIC)在F1和F2代中分别为:3.2,0.528,0.463和3.2,0.496,0.447。结果表明大多数微卫星标记位点表现为中高度杂合性。在资源群体中,平均有信息减数分裂数是217.4(44-316),而各染色体上两性平均图谱的长度分别是:172.3cM(SSC4),168.7cM(SSC6),191.7cM(SSC7),197.3cM(SSC8),178.3cM(SSC13)。与USDA-MARC的参考图谱相比,标记位点的顺序相同,但长度均较长。雌雄两性图谱相比,第4和第6染色体上雌性图谱长于雄性图谱;而在另外3条染色体上,则雄性图谱长于雌性图谱。结果显示了标记位点在资源猪群的遗传特征和遗传关系,其连锁图谱可用于今后的QTL定位。     

柑桔BES SSR标记开发及连锁图延伸和加密

为了系统性地开发和拓展柑桔SSR标记,通过对公布的柑桔BAC文库末端序列(BAC-End sequence,BES)进行SSR分析,选择1500个SSR位点设计合成并检测323对引物。结果表明:(1)从总长度为28.1 Mb的46 339条序列中共检测出22 403个SSR位点,约每2条序列就会出现一个SSR位点,发生频率为48%,相当于平均1.25kb的序列中就会出现1个SSR,频率约为柑桔EST的2倍,且不同核心重复序列的SSR发生特点与EST也不同。(2)所合成的323对引物中,有效扩增316对,扩增率约98%,173对表现多态性,总多态性比率约55%,多态性引物中单核苷酸重复类型15对,双核苷酸重复类型100对,三核苷酸及以上重复类型58对,表明柑桔BES中具有较为丰富的多态性SSR标记。(3)结合已发表的遗传作图数据,对总计349个多态性位点进行遗传连锁分析,获得的新遗传连锁图谱共含有9个连锁群、334个SSR标记、总长844.2cM、平均图距2.53cM,延长和加密了先前的图谱。该研究开发的新SSR标记为开展柑桔遗传鉴定分析和遗传图谱构建提供了新的标记来源,加密的遗传图谱为柑桔的基因定位、图位克隆和标记辅助育种等奠定了基础,SSR分析结果也为其他物种SSR标记的开发提供了参考。     

Integration of the Physical and Genetic Linkage Map for Human Chromosome 13

We have used a panel of somatic cell hybrids containing different rearrangements of human chromosome 13 to integrate genetic and physical maps of this chromosome. The positions of 17 translocation/deletion breakpoints on human chromosome 13 have been determined relative to the microsatellite markers on the genetic linkage map compiled by Généthon. Because markers on maps from several other Consortium groups have also been analyzed using many of the same hybrids, it was possible to relate these with the Généthon map. The position of all of the chromosome breakpoints have been placed, wherever possible, between two adjacent markers on the genetic linkage maps using PCR analysis for the presence/absence of the markers in the somatic cell hybrids. The positions of the breakpoints have already been determined cytogenetically, and some of these breakpoints are located at landmark positions on the chromosome. The relative density of markers along the chromosome differs between independently derived maps, and, based on the known locations of certain breakpoints in the physical map, inconsistencies in the genetic maps have been identified.     

RAPD和SSR两种标记构建的中国对虾遗传连锁图谱

利用RAPD和SSR分子标记结合拟测交策略,对中国对虾(Fenneropenaeuschinensis)“黄海1号”雌虾与野生雄虾作为亲本进行单对杂交产生的F1代,采用RAPD和SSR两种分子标记技术初步构建了中国对虾雌、雄遗传连锁图谱。对460个RAPD引物和44对SSR引物进行筛选,共选出61个RAPD引物和20对SSR引物,用于对父母本和82个F1个体进行遗传分析。共得到母本分离标记146个(RAPD标记128个,微卫星标记18个)和父本分离标记127个(RAPD标记109个,微卫星标记18个)。雌性图谱包括8个连锁群、9个三联体和14个连锁对,标记间平均间隔为11·28cM,图谱共覆盖1173cM,覆盖率为59·36%;雄性图谱包括10个连锁群、12个三联体和7个连锁对,标记间平均间隔为12·05cM,图谱共覆盖1144·6cM,覆盖率为62·01%。中国对虾遗传图谱的构建为其分子标记辅助育种、比较基因组作图及数量性状位点的定位与克隆奠定了基础。     

用微卫星标记构建两系稻培矮64s/E32的分子遗传连锁图 Construction of a Microsatellite Linkage Map in a DH Population

用两系杂交稻强优组合培矮64s/E32的一个加倍单倍体（DH）群体,共86个株系,构建了水稻的SSR标记遗传连锁图。选用美国康耐尔大学公布的302对SSR引物,共有127对在两个亲本间检测到多态性,比率为4205%。建成的水稻染色体的图谱（记为PEMAP）共包含122个SSLP标记座位,总长度为1213.4 cM。PEMAP与Temnykh等发表的图谱（记为CUMAP）具有很高的可比性,绝大多数标记都被定位于相同的染色体上,且排列顺序一致。该DH群体的偏分离情况较严重,122个标记座位中有34个发生显著偏分离,比例达27.8%。值得注意的是,在第1、3、10、11染色体上的标记全部偏向培矮64s,第4、6、7、8、9染色体上的标记则全部偏向E32。 Abstract:A doubled haploid population (DH) consisting of 86 lines derived by anther culture of Peiai64s/E32,a two-line hybrid rice variety with high heterosis,was used to construct a microsatellite or SSLP linkage map of rice chromosomes.A total of 302 PCR primers for SSLP analysis on these chromosomes were chosen from a map published by Cornell University (designated CUMAP) and 127 (42.05%) of them were found polymorphic between the two parents.Those polymorphic PCR primers were used for population genotyping.The map (designated PEMAP) comprises 122 microsatellite maker loci,covering a total length of 1213.4 cM.The PEMAP is highly comparable with the CUMAP.Most of the markers were mapped onto the same chromosomes and aligned in the same order.Serious segregation distortion was observed in this DH population,with 34 (27.8%) markers showing significant deviation.It is noted that all markers on chromosomes 1,3,10 and 11 were biased to Peiai64s,while those on chromosomes 4,6,7,8 and 9 were opposite.     

Use of Random Amplified Polymorphic DNA Markers for Mapping the Chickpea Genome

Three interspecific crosses were developed using Cicer arietinum (ICC 4918) as the female parent and wild Cicer species [C. reticulatum - JM 2100, JM 2106 and C. echinospermum - ICCW 44] as the male parent. Cicer arietinum (ICC 4918) × C. reticulatum (JM 2100) cross produced the largest number of F₂ plants and was chosen for linkage mapping using Random Amplified Polymorphic DNA (RAPD) primers. A partial linkage map was constructed based upon the segregation of 36 RAPD markers obtained by amplification using 35 primers. The linkage map consists of two linkage groups with 17 linked markers covering a total of 464.9 cM. Analyses also revealed association of three morphological traits with linked RAPD markers. Out of seven morphological traits tested for association with linked markers in the segregating plants, four Quantitative trait loci (QTL) were detected for the trait leaf length and three QTLs each for the traits leaf width and erect plant habit.     

家蚕分子连锁图谱的构建及分子标记育种研究进展

国际蚕分子育种计划（ＩｎｔｅｒｎａｔｉｏｎａｌＳｉｌｋｗｏｒｍＰｒｏｊｅｃｔ）是ＲｈｏｄｅＩｓｌａｎｄ大学的Ｍ．Ｒ．Ｇｏｌｄｓｍｉｔｈ在１９９１年提出来的,也叫基因标记育种计划〔１〕。该计划主要包括两步工作：第一步是制作蚕的分子基因图,第二步是数量性状定位分析（简称ＱＴＬ分析,也叫数量性状定位作图：ＱＴＬＭａｐｐｉｎｇ）。最后利用分子标记直接在ＤＮＡ水平进行重要经济性状（数量性状）的选择、固定,即实施“分子育种”,用很短时间育成兼具高抗、强健、丝多、质优、易繁等特性的新蚕品种,本文对此进行简要介…     

家蚕分子连锁图谱的构建及分子标记育种研究进展

国际蚕分子育种计划(International Silkworm Project)是Rhode Island大学的M.R.Goldsmith在1991年提出来的,也叫基因标记育种计划[1].该计划主要包括两步工作:第一步是制作蚕的分子基因图, 第二步是数量性状定位分析(简称QTL分析,也叫数量性状定位作图:QTL Mapping).最后利用分子标记直接在DNA水平进行重要经济性状(数量性状)的选择、固定,即实施"分子育种",用很短时间育成兼具高抗、强健、丝多、质优、易繁等特性的新蚕品种,本文对此进行简要介绍.     

用AFLP标记快速构建遗传连锁图谱并定位一个新基因tms5

报导了一个分子标记连锁图的快速构建方法.通过对水稻(Oryza sativa L.)"安农S-1"和"南京11"的F2分离群体的AFLP分析找到了142个AFLP标记,用这142个AFLP标记以及已定位的25个SSR标记和5个RFLP标记构建了水稻12个染色体的分子标记连锁图,该图覆盖水稻基因组的1 537.4 cM,相邻标记间的平均间距为9.0 cM,这是在国内建立的第一张AFLP标记连锁图.在建立连锁图谱的同时把一个新基因tms5 (水稻温敏核不育基因)定位在第2染色体上.     

用AFLP标记快速构建遗传连锁图谱并定位一个新基因tms5

报导了一个分子标记连锁图的快速构建方法。通过对水稻(Oryza sativa L．)“安农S-1”和“南京11”的F2分离群体的AFLP分析找到了142个AFLP标记,用这142个AFLP标记以及已定位的25个SSR标记和5个RFIP标记构建了水稻12个染色体的分子标记连锁图,该图覆盖水稻基因组的1537．4cM,相邻标记间的平均间距为9．0cM,这是在国内建立的第一张AFLP标记连锁图。在建立连锁图谱的同时把一个新基因tms5(水稻温敏核不育基因)定位在第2染色体上。     

A Primary Genetic Linkage Map of 14 Polymorphic Loci for the Short Arm of Human Chromosome 8

A genetic linkage map of markers for the short arm of human chromosome 8 has been constructed with 14 polymorphic DNA markers on the basis of genotypes obtained in 40 CEPH reference families. This unbroken map spans 45 cM in males and 79 cM in females. The 14 markers include three genes, MSR, LPL, and NEFL, and one anonymous DNA segment that were previously assigned to chromosome 8. The other 10 marker had been isolated from a chromosome 8-specific cosmid library and physically localized to chromosomal bands by fluorescence in situ hybridization. The order of loci determined by genetic linkage was consistent with their physical locations. This map will facilitate efficient linkage studies of human genetic diseases that may be segregating on chromosome 8p and will provide anchor points for development of high-resolution maps for this chromosomal region.     

利用两个测序水稻品种构建微卫星连锁图谱

利用已完成基因组测序的两个水稻品种日本晴和931l的数据库成功开发出水稻微卫星新标记,并利用由90个单株组成的日本晴×9311 F2作图群体,构建了一张包含152个SSR标记位点、覆盖基因组总长度2 455.7 cM的连锁图谱,有46个SSR新标记为自主开发,该图谱标记间的平均遗传距离为16.16 cM;并将未能在Temnykh等人(2001)构建的图谱上定位的微卫星标记RM345和RM494定位在第6染色体上.通过与Temnykh等人(2001)和兰涛等人(2003)所构建的图谱从作图群体的类型和大小、标记的类型和数量、标记在染色体上的线性排列顺序等几个方面进行比较,所绘制的图谱其标记在染色体线性排列上与Temnykh等人绘制的图谱具有很高的一致性,达93.81%.