High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones

Cosmid clones containing human DNA inserts have been mapped on chromosome 11 by fluorescence in situ hybridization under conditions that suppress signal from repetitive DNA sequences. Thirteen known genes, one chromosome 11-specific DNA repeat, and 36 random clones were analyzed. High-resolution mapping was facilitated by using digital imaging microscopy and by analyzing extended (prometaphase) chromosomes. The map coordinates established by in situ hybridization showed a one to one correspondence with those determined by Southern (DNA) blot analysis of hybrid cell lines containing fragments of chromosome 11. Furthermore, by hybridizing three or more cosmids simultaneously, gene order on the chromosome could be established unequivocally. These results demonstrate the feasibility of rapidly producing high-resolution maps of human chromosomes by in situ hybridization.     

A physical map of the 1-gigabase bread wheat chromosome 3B

As the staple food for 35% of the world's population, wheat is one of the most important crop species. To date, sequence-based tools to accelerate wheat improvement are lacking. As part of the international effort to sequence the 17-billion-base-pair hexaploid bread wheat genome (2n = 6x = 42 chromosomes), we constructed a bacterial artificial chromosome (BAC)-based integrated physical map of the largest chromosome, 3B, that alone is 995 megabases. A chromosome-specific BAC library was used to assemble 82% of the chromosome into 1036 contigs that were anchored with 1443 molecular markers, providing a major resource for genetic and genomic studies. This physical map establishes a template for the remaining wheat chromosomes and demonstrates the feasibility of constructing physical maps in large, complex, polyploid genomes with a chromosome-based approach.     

Megabase-scale mapping of the HLA gene complex by pulsed field gel electrophoresis

In the study of the genetic structure of mammalian chromosomes, there exists a "resolution gap" between molecular cloning experiments and meiotic linkage analyses. This gap has discouraged attempts to construct full-scale genetic maps of mammalian chromosomes. The organization of the human major histocompatibility complex was examined within this range by pulsed field gel electrophoresis. The data obtained indicate that the complex spans over 3000 kilobases and enable the construction of a megabase-scale molecular map. These results indicate that the techniques employed in DNA extraction, enzymatic digestion, electrophoresis, and hybridization are suitable for the efficient analysis of megabase regions of mammalian chromosomes and effectively bridge the resolution gap between molecular cloning and classical genetics.     

Chromosomal region of the cystic fibrosis gene in yeast artificial chromosomes: a model for human genome mapping

A general strategy for cloning and mapping large regions of human DNA with yeast artificial chromosomes (YAC's) is described. It relies on the use of the polymerase chain reaction to detect DNA landmarks called sequence-tagged sites (STS's) within YAC clones. The method was applied to the region of human chromosome 7 containing the cystic fibrosis (CF) gene. Thirty YAC clones from this region were analyzed, and a contig map that spans more than 1,500,000 base pairs was assembled. Individual YAC's as large as 790 kilobase pairs and containing the entire CF gene were constructed in vivo by meiotic recombination in yeast between pairs of overlapping YAC's.     

Polymorphism analysis of microsatellites and construction of linkage map in part regions of four chromosomes in chicken

Based on chicken' consensus map issued in 2000,17 microsatellites near 4 candidate genes such as IGF2,OBR,GDF8 and APOA1 in 4 chromosomes(chromosome 5,7,8 and 24)were chosen for polymorphism analysis and construction of linkage map.Combining the technique of PCR and the fluorescent semi-automated detection,genome scanning was performed for 440 chickens,which was derived from China Agricultural University chicken resource families within three generations.The individuals of this resource families were genotyped.The results showed that the number of alleles ranged from 4 to 14;heterozygosity(H) of markers was between 0.3116 and 0.9148.Polymorphic information content(PIC)varied from 0.2672 to 0.8679.Microsatellites along with above-mentioned 4 candidate genes doing as general markers were used to construct linkage map.The spans of 4 linkage maps constructed in the part region of chromosome 5,7,8 and 24 were 263.5,79.9,206.2 and 104.2 cM,respectively.The order of markers was consistent with that of counterpart of reported consensus map.However,The spans of linkage map were larger than that of consensus map.The constructed linkage maps laid the foundation for mapping quantitative trait loci(QTL)responsible for economically important traits in chicken.     

鲜食甜玉米南方锈病抗性QTL定位分析

为挖掘新的抗南方锈病基因资源,本研究以甜玉米组合M5×M114的216个F2单株为遗传作图群体,应用BSA方法从500对SSR引物中筛选出2对在F2代抗病和感病DNA池间具有多态性的引物,分别位于4和9号染色体上;在4和9号染色体上重新设计100对SSR引物,构建了包含33个标记位点总长为241.2cM的连锁遗传图,各个标记间的平均距离为7.53cM。结合F2单株对南方锈病的抗性表现,用复合区间作图法在4和9号染色体上共检测到7个显著的南方锈病抗性QTLs,其中:4个QTLs位于4号染色体上,可解释12.1%、7.8%、18.2%和14.9%表型变异;3个位于9号染色体上,分别解释17.0%、13.3%与19.2%的表型变异。研究结果可为抗南方锈病的精细定位、主效基因克隆和抗南方锈病鲜食甜玉米品种选育提供理论依据。     

A BAC-based physical map of the major autosomes of Drosophila melanogaster

We constructed a bacterial artificial chromosome (BAC)-based physical map of chromosomes 2 and 3 of Drosophila melanogaster, which constitute 81% of the genome. Sequence tagged site (STS) content, restriction fingerprinting, and polytene chromosome in situ hybridization approaches were integrated to produce a map spanning the euchromatin. Three of five remaining gaps are in repeat-rich regions near the centromeres. A tiling path of clones spanning this map and STS maps of chromosomes X and 4 was sequenced to low coverage; the maps and tiling path sequence were used to support and verify the whole-genome sequence assembly, and tiling path BACs were used as templates in sequence finishing.     

The genetic linkage map of the human X chromosome

A database useful for mapping the human X chromosome has been established. The data consist of the genotypic characterizations obtained at more than 20 DNA marker loci from a set of 38 selected families. Multilocus linkage analysis has provided an initial genetic map completely spanning the distance from the distal short arm to the distal long arm of the chromosome, for a total genetic length of at least 185 recombination units. Analysis of the recombinational behavior of fully marked chromosomes suggests that the number of recombination events on the X chromosome may be nonrandom. Linkage studies of six families that carry the mutation which causes Duchenne muscular dystrophy were combined with linkage data from a large number of normal families. This permitted mapping of the locus for Duchenne muscular dystrophy with greater precision and statistical confidence than studies in which disease families alone provided the genotypic database. This observation suggests that the normal linkage map of this chromosome should be especially valuable in the mapping of rare X-linked diseases.     

水稻RFLP遗传作图群体重建的初步研究

用已定位的94个RFLP标记为探针,测定了12种限制性内切酶酶切后的F_2群体中58个单株及其双亲的RFLP表现。根据各标记F_2群体中的连锁分析资料,初步重建了水稻RFLP遗传作图群体。结果表明:51个标记被定位在10条染色体上,它们的连锁群和定位位置相同于原遗传图;18个标记揭示了双亲间的多态性,但是,没有发现与其它标记的连锁关系;12个标记至少在1种酶切条件下揭示出RFLP,然而,它们现属的连锁群有别于原遗传图;其余13个标记无法揭示RFLP存在。这一遗传群体可供新的DNA标记的定位。     

一种基于聚合酶链式反应检测SNP的方法

SNP是具有广泛利用潜力的第3代分子标记,本文旨在开发一种利用PCR技术快速检测SNP的方法。设计思路是:根据已知SNP位点设计2条特异正向引物,其最后一个碱基分别与已知SNP的2个碱基相同,同时在1条引物的5′端添加1段20 bp左右的其他物种的特异序列(如细菌DNA序列),然后选择1条合适的反向引物;最后同时加入3条引物,通过梯度PCR选择合适的退火温度进行PCR反应。利用这一方法成功将玉米的ZDS基因定位在玉米第7染色体短臂7.02 Bin。这种检测SNP的方法设计简单,费用低廉,尤其适合SNP标记的分子标记连锁图构建或者基因定位。     

生物技术与葡萄遗传育种

 生物技术的发展和应用已渗透到葡萄遗传育种的各个领域。DNA分子标记为研究葡萄品种起源和育成新品种提供了强有力的工具,特别是SSR标记在葡萄品种鉴别、系谱分析和遗传图谱构建方面已得到广泛应用。基于DNA分子标记遗传图的构建加速了杂种的早期选择,遗传图和物理图的结合可以克隆控制重要性状的功能基因,遗传转化技术的应用可以改良现有的葡萄品种。因此,生物技术正深刻改变着传统的葡萄育种方法。     

A genetic linkage map with 178 SSR and 1901 SNP markers constructed using a RIL population in wheat (Triticum aestivum L.)

The construction of high density genetic linkage map provides a powerful tool to detect and map quantitative trait loci(QTLs) controlling agronomically important traits. In this study, simple sequence repeat(SSR) markers and Illumina 9K i Select single nucleotide polymorphism(SNP) genechip were employed to construct one genetic linkage map of common wheat(Triticum aestivum L.) using 191 recombinant inbred lines(RILs) derived from cross Yu 8679×Jing 411. This map included 1 901 SNP loci and 178 SSR loci, covering 1 659.9 c M and 1 000 marker bins, with an average interval distance of 1.66 c M. A, B and D genomes covered 719.1, 703.5 and 237.3 c M, with an average interval distance of 1.66, 1.45 and 2.9 c M, respectively. Notably, the genetic linkage map covered 20 chromosomes, with the exception of chromosome 5D. Bioinformatics analysis revealed that 1 754(92.27%) of 1 901 mapped SNP loci could be aligned to 1 215 distinct wheat unigenes, among which 1 184(97.4%) were located on o ne single chromosome, and the rest 31(2.6%) were located on 2 to 3 chromosomes. By performing in silico comparison, 214 chromosome deletion bin-mapped expressed sequence tags(ESTs), 1 043 Brachypodium genes and 1 033 rice genes were further added onto the genetic linkage map. This map not only integrated genetic and physical maps, SSR and SNP loci, respectively, but also provided the information of Brachypodium and rice genes corresponding to 1 754 SNP loci. Therefore, it will be a useful tool for comparative genomics analysis, fine mapping of QTL/gene controlling agronomically important traits and marker-assisted selection breeding in wheat.     

水稻籼粳交F8、F2群体穗长QTL比较分析

【目的】对水稻F8重组自交系群体穗长QTL进行检测,并比较分析相同亲本衍生的不同群体的遗传图谱、QTL位置、QTL效应的异同,鉴定稳定表达的穗长QTL,以期增加对穗长遗传行为的了解,且有助于通过分子聚合育种手段改良穗长性状。【方法】以籼稻品种泸恢99和粳稻品种日本晴（基因组测序）为亲本构建的F8重组自交系群体中的188个家系为研究材料,利用包含207个标记的遗传连锁图谱,采用基于混合线性模型的QTL定位软件QTLNetwork 2.0,对水稻穗长QTL进行定位和效应分析,并比较分析F8、F2群体的QTL定位和遗传图谱异同。【结果】在F8群体中检测到7个与穗长性状相关的QTL,分别位于第2、3、6、7、8、10染色体上,QTL对表型变异的贡献率为3.38%—14.8%,总贡献率为52.5%。F8、F2群体在5条相同染色体上都定位到了穗长QTL,这些QTL所在标记区间物理位置大部分是重叠和包含关系。F8、F2图谱在定位标记数、标记的位置顺序、遗传距离、平均图距等方面发生了变化。【结论】在F8、F2群体检测到一个稳定遗传的主效应QTL位点,位于第6染色体,并发现了4个尚未报道的穗长QTL。     

糯玉米耐贮藏相关性状的主效QTL定位

为挖掘、利用玉米耐贮藏基因资源,对糯玉米组合N7×N32的216个F2单株种子进行2年的常温贮藏,并以贮藏后种子的单位质量电导率和发芽率为耐贮藏性指标,建立极端耐与不耐贮藏BSA混池.首先筛选在2个BSA混池间有多态性的引物,后在多态引物所在的染色体上增加引物,以F2为作图群体,构建分子标记连锁图谱;同时结合测得的F2...     

兰科植物遗传图谱与QTL定位研究进展

兰科植物俗称兰花,是最重要的观赏花卉,构建遗传图谱,特别是高密度遗传图谱有助于提高育种水平,本研究主要对兰科植物遗传图谱与重要性状的QTL定位进行了综述。结果表明:1)2007年报道出兰科石斛属第一张遗传图谱以来,累计发表了13张兰科植物遗传连锁图谱。2)从图谱类型来看,5张为品种间图谱,8张为种间图谱;从图谱用途来分,7张可以用于QTL定位,3张可用于精细定位或基因克隆,另3张为基础参考图谱。3)从已构建的兰科植物遗传图谱看,总图距越来越高且平均图距越来越小,为图谱的实际应用奠定了基础,但作图群体较小始终是个问题。4)虽然在蝴蝶兰叶片、花色,石斛兰萼片大小及石斛茎及多糖含量等相关性状上进行了QTL定位,但在花朵大小、花朵数、花香、花型、花斑和植株抗性等方面均未涉及。因此,构建遗传图谱,特别是高密度遗传图谱能够为兰花的生物学研究及分子标记辅助育种提供参考依据。     

Assignment of the gene for myelin proteolipid protein to the X chromosome: implications for X-linked myelin disorders

Several inherited disorders in humans and in rodents result in myelin dysgenesis and a deficiency of the molecular constituents of myelin. A complementary DNA to one of the two major myelin proteins, myelin proteolipid protein (also known as lipophilin), has been used with Southern blot analysis of somatic cell hybrid DNA to map the human proteolipid protein gene to the middle of the long arm of the human X chromosome (bands Xq13-Xq22) and to assign the murine proteolipid protein gene to the mouse X chromosome. Comparison of the gene maps of the human and mouse X chromosomes suggests that myelin proteolipid protein may be involved in X-linked mutations at the mouse jimpy locus and has implications for Pelizaeus-Merzbacher disease, a human inherited X-linked myelin disorder.     

用2个不同来源的DNA探针构建水稻RFLP连锁图谱(英文)

基于籼稻品种圭 6 30与粳稻品种台湾粳杂交产生的一个包含 111个株系的加倍单倍体 (DH)群体 ,构建了一个水稻 RFL P连锁图谱 .该图谱 (记为 DH图谱 )含有 175个 RFL P座位 ,全长 12 2 4 .6 c M,相邻标记间平均距离 7.0 c M.用于 RFL P分析的探针选自分别由日本水稻基因组计划 (RGP)和美国康耐尔大学 (CU)构建的 2个水稻图谱 .9个染色体区域发现明显的偏分离 ,但 DH图谱与 RGP和 CU图谱之间仍表现出较好的共线性 .因此 ,DH图谱可以作为那 2张图谱之间遗传信息相互沟通的桥梁 .本图谱还可用于对水稻中控制重要农艺性状的数量性状基因座的定位和分析     

甜瓜SLAF图谱构建及果实相关性状QTL分析

【目的】通过对甜瓜果实相关性状进行QTL定位及候选基因分析,为甜瓜品质育种及基因挖掘与功能验证提供理论基础。【方法】利用薄皮甜瓜1244为母本、厚皮甜瓜M5为父本配置杂交组合,结合SFLA测序技术开发分子标签,构建高密度遗传图谱,以F_2:3表型数据为基础,采用Mapqtl进行QTL定位分析。【结果】获得了380 446 Mreads（83.12 Gb）数据,测序平均Q30为93.59%,平均GC含量为36.87%;开发了112 844个SLAF标签,3 274 879个SNP;构建了12个连锁群,共计10 596个上图标记,总图距为1 383.88 cM,标记间平均距离为0.13 cM,上图标记完整度平均为99.92%。将控制甜瓜果面沟性状基因（fst）定位在第11条染色体末端Marker 1993423（62.18）—Marker 1998820（63.05）,覆盖基因组0.72 Mb,包含33个候选基因;控制甜瓜果皮花纹基因（fpp）定位在第2条染色体Marker 459584（90.91）—Marker 459446（90.91）,覆盖基因组0.08 Mb,包含5个候选基因,其中MELO3C026292（1-脱氧-D-木酮糖-5-磷酸还原异构酶）可能为控制果皮花纹的候选基因;同时检测到甜瓜果皮底色1个QTL位点fpc,位于第7条染色体Marker 1229174（7.14）—Marker 1229973（7.14）,贡献率为9.9%;检测到果型指数1个QTL位点fs9.1,位于第9条染色体Marker 1705671（76.19）—Marker 1705915（79.23）,贡献率为7.6%;在第1、2、6、7、10染色体检测到单果重相关6个QTL位点（sfw1.1、sfw2.1、sfw2.2、sfw6.1、sfw7.1、sfw10.1）,贡献率在3.1%—17.0%,LOD值介于3.0—5.6。【结论】将甜瓜果面沟、果皮花纹基因分别定位在第11和第2条染色体,分别获得33个和5个候选基因;检测到1个果皮底色QTL位点、1个果型指数QTL位点和6个单果重QTL位点。     

鲤45S rDNA的染色体荧光原位杂交定位

应用荧光原位杂交技术,通过设计位于5.8S rDNA、18S rDNA和非转录IGS区域的3条探针CAAG1191、CAAG1845和CAAG3602,分别对散鳞镜鲤Cyprinus carpio var.scattered mirror和松浦鲤Cyprinus carpio Songpu的45S核糖体DNA(ribosomal DNA,rDNA)进行染色体定位及共定位.结果表明:45S rDNA均位于两品种鲤一对近端着丝粒染色体的短臂末端,具有染色体特异性,表明45S rDNA序列的探针能够在鲤细胞遗传学研究中用于标识其所在染色体,并与鲤遗传连锁图谱中长度为227 cM的1号连锁群相对应;两品种鲤的染色体数目均为2n=100,45S rDNA在鲤基因组内仅定位于一对同源染色体,不存在复制位点,证实了鲤基因组在全基因组复制事件之后又经历了重新二倍化过程.