用近等基因池法定位与水稻柱头外露率相关的QTL

利用水稻柱头外露率差异极显著的亲本Ⅱ-32B和冈46B杂交,对亲本、F1和F2群体进行柱头外露率鉴定,F1柱头外露率介于高、低柱头外露率亲本Ⅱ-32B和冈46B之间;F2群体柱头外露率表现正态分布,说明柱头外露率是由多基因控制的数量性状.在498个F2单株群体中选择极端高柱头外露率和极端低柱头外露率的单株分别组成高、低柱头外露率池,利用914对SSR引物对其进行PCR扩增多态性检测结果发现,引物RM3437、RM31、RM3188、RM5310和RM3395在两亲本和两池间均检测到多态性;单标记分析法表明,第5染色体上的RM3437和RM31,第2染色体上的RM3188,第1染色体上的RM5310和第8染色体上的RM3395与柱头外露率呈极显著连锁关系,对该群体柱头外露率的贡献率分别为13.65%、13.62%、7.09%、4.63%和4.83%.第5染色体上的RM3437、RM31可能与同一个主效QTL连锁,该两标记可用于水稻高柱头外露率不育系的分子标记辅助选择育种.     

The effect of histone gene deletions on chromatin structure in Saccharomyces cerevisiae

As a way of studying nucleosome assembly and maintenance in Saccharomyces cerevisiae, mutants bearing deletions or duplications of the genes encoding histones H2A and H2B were analyzed. Previous genetic analysis had shown that only one of these mutants exhibited dramatic and pleiotropic phenotypes. This mutant was also the only one that contained disrupted chromatin, suggesting that the original phenotypes were attributable to alterations in chromosome structure. The chromatin disruption in the mutant, however, did not extend over the entire genome, but rather was localized to specific regions. Thus, while the arrangement of nucleosomes over the HIS4 and GAL1 genes, the telomeres, and the long terminal repeats (delta sequences) of Ty retrotransposons appeared essentially normal, nucleosomes over the CYH2 and UBI4 genes and the centromere of chromosome III were dramatically disrupted. The observation that the mutant exhibited localized chromatin disruptions implies that the assembly or maintenance of nucleosomes differs over different parts of the yeast genome.     

Human phosphoglycerate kinase and inactivation of the X chromosome

The fibroblasts derived from the skin of a woman heterozygous for an X-linked deficiency of phosphoglycerate kinase represented a mosaic. Two of 22 clones with normal glucose-6-phosphate dehydrogenase activity and hypoxanthine(guanine) phosphoribosyltransferase activity had no phosphoglycerate kinase activity detected by electrophoresis. Because the loci for glucose-6-phosphate dehydrogeniase and hypoxanthine(guanine)phosphoribosyltransferase are already known to undergo inactivation and to be on the short arm of the X chromosome and the locus for phosphoglycerate kinase is on the long arm, these observations support the conclusion that the entire human X chromosome can be involved in X inactivation.     

Capturing chromosome conformation

We describe an approach to detect the frequency of interaction between any two genomic loci. Generation of a matrix of interaction frequencies between sites on the same or different chromosomes reveals their relative spatial disposition and provides information about the physical properties of the chromatin fiber. This methodology can be applied to the spatial organization of entire genomes in organisms from bacteria to human. Using the yeast Saccharomyces cerevisiae, we could confirm known qualitative features of chromosome organization within the nucleus and dynamic changes in that organization during meiosis. We also analyzed yeast chromosome III at the G1 stage of the cell cycle. We found that chromatin is highly flexible throughout. Furthermore, functionally distinct AT- and GC-rich domains were found to exhibit different conformations, and a population-average 3D model of chromosome III could be determined. Chromosome III emerges as a contorted ring.     

Recruitment and spreading of the C. elegans dosage compensation complex along X chromosomes

To achieve X-chromosome dosage compensation, organisms must distinguish X chromosomes from autosomes. We identified multiple, cis-acting regions that recruit the Caenorhabditis elegans dosage compensation complex (DCC) through a search for regions of X that bind the complex when detached from X. The DCC normally assembles along the entire X chromosome, but not all detached regions recruit the complex, despite having genes known to be dosage compensated on the native X. Thus, the DCC binds first to recruitment sites, then spreads to neighboring X regions to accomplish chromosome-wide gene repression. From a large chromosomal domain, we defined a 793-base pair fragment that functions in vivo as an X-recognition element to recruit the DCC.     

Isolation of single-copy human genes from a library of yeast artificial chromosome clones

A recently developed cloning system based on the propagation of large DNA molecules as linear, artificial chromosomes in the yeast Saccharomyces cerevisiae provides a potential method of cloning the entire human genome in segments of several hundred kilobase pairs. Most application of this system will require the ability to recover specific sequences from libraries of yeast artificial chromosome clones and to propagate these sequences in yeast without alterations. Two single-copy genes have now been cloned from a library of yeast artificial chromosome clones that was prepared from total human DNA. Multiple, independent isolates were obtained of the genes encoding factor IX and plasminogen activator inhibitor type 2. The clones, which ranged in size from 60 to 650 kilobases, were stable on prolonged propagation in yeast and appear to contain faithful replicas of human DNA.     

Distinct cohesin complexes organize meiotic chromosome domains

Meiotic cohesin complexes at centromeres behave differently from those along chromosome arms, but the basis for these differences has remained elusive. The fission yeast cohesin molecule Rec8 largely replaces its mitotic counterpart, Rad21/Scc1, along the entire chromosome during meiosis. Here we show that Rec8 complexes along chromosome arms contain Rec11, whereas those in the vicinity of centromeres have a different partner subunit, Psc3. The arm associated Rec8-Rec11 complexes are critical for meiotic recombination. The Rec8-Psc3 complexes comprise two different types of assemblies. First, pericentromeric Rec8-Psc3 complexes depend on histone methylation-directed heterochromatin for their localization and are required for cohesion during meiosis II. Second, central core Rec8-Psc3 complexes form independently of heterochromatin and are presumably required for establishing monopolar attachment at meiosis I. These findings define distinct modes of assembly and functions for cohesin complexes at different regions along chromosomes.     

Site-specific cleavage of human chromosome 4 mediated by triple-helix formation.

Direct physical isolation of specific DNA segments from the human genome is a necessary goal in human genetics. For testing whether triple-helix mediated enzymatic cleavage can liberate a specific segment of a human chromosome, the tip of human chromosome 4, which contains the entire candidate region for the Huntington's disease gene, was chosen as a target. A 16-base pyrimidine oligodeoxyribonucleotide was able to locate a 16-base pair purine target site within more than 10 gigabase pairs of genomic DNA and mediate the exact enzymatic cleavage at that site in more than 80 percent yield. The recognition motif is sufficiently generalizable that most cosmids should contain a sequence targetable by triple-helix formation. This method may facilitate the orchestrated dissection of human chromosomes from normal and affected individuals into megabase sized fragments and facilitate the isolation of candidate gene loci.     

利用TAC系统构建多个光合同化物运输蛋白基因转化载体和水稻的遗传转化

采用人工染色体(TAC)多基因组装载体系统,将一组与水稻光合同化物运输转化有关基因构建在同一载体上,这些基因分别是蔗糖转运蛋白基因(OsSUT5),单糖转运蛋白(OsMST4)、细胞壁转化酶(OsCIN3)和ATP/ADP转运蛋白(OAAT),获得了包含这4个基因的植物表达载体pTAC747H-OAAT-SUT5-CIN3-MST4,质粒大小近30 kb。通过农杆菌转化法,将基因导入水稻,以探讨这些基因对水稻光合同化物运输和转化的调控作用。     

普通小麦-华山新麦草异附加系的SSR分析

为了建立一套完整的小麦-华山新麦草异附加系,从筛选出的46对在华山新麦草与小麦间具有多态性的SSR引物中选出7对扩增条带清晰的引物,对20个小麦-华山新麦草二体附加系进行归类分析。结果发现:出现了11个小麦-华山新麦草异附加系类型,说明在这20个附加系中包括了华山新麦草7个同源群的附加系,并探讨了小麦与华山新麦草杂交及其后代衍生过程中发生华山新麦草染色体间重排的可能。     

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.     

果蝇(Drosophila melanogaster)唾腺染色体的制备及染色体识别

果蝇(Drosophila melanogaster)唾腺染色体的制备与分析是果蝇细胞遗传研究的基本实验技术,也是非常流行的遗传学教学实验。国内绝大多数的实验教材介绍了这个实验的操作方法,但对于如何辨别染色体各条臂,并进而识别具体的带纹均未予以介绍。笔者详细介绍唾腺染色体的制备及拍摄、数码照片拼接方法;并着重介绍如何根据各条臂端粒端的形态特征,以及利用Lefevre的唾腺染色体照片图谱中指示的蓬突(puff)、缢缩及带纹组合特征识别染色体臂的方法,供研究和实验教学参考。     

普通小麦与华山新麦草、簇毛麦杂交后代染色体形态观察

对普通小麦(Tritcium aestivum)与华山新麦草(Psathyrostachys huashanica)杂交后代H9802-6、普通小麦与簇毛麦(Haynaldia villosa)杂交后代V9910-15-4花粉的减数分裂进行了细胞学观察。结果表明:H9802-6出现了异代换系和异附加系,因此该材料还不是很稳定;V9910-15-4后代个体中染色体数目、染色体构型多样复杂,染色体联会过程中出现了环状联会、顶端联会、单价体不联会等异常情况,其杂交后代的稳定及利用较困难。     

Nonrandomness of translocations in man: preferential entry of chromosomes into 13-15-21 translocations

Lymphocytes from 20 individuals with Down's syndrome due to 13-15/21 centric-fusion translocations were studied by autoradiography after continuous late labeling with tritiated thymidine. In no case was chromosome 13 involved; chromosome 14 was involved in 18 cases, and chromosome 15 in two cases. These results are similar to those from 13 previously studied cases and indicate that the entry of chromosomes 13-15 into translocations is nonrandom. This nonrandomness is not a simple function of chromosome size or shape, since chromosomes 13-15 are acrocentrics of similar size.     

陕西大豆初选核心种质的代表性分析

比较了1 035份陕西大豆种质与从中选出的102份初选核心种质间15个农艺性状的差异,检验了初选核心种质的代表性。结果表明,初选核心种质的生育类型、种皮色、生长习性等15个农艺性状表型频率和生育期、百粒重、株高等5个数量性状的平均数、标准差、变异系数等指标与总体资源基本一致;初选核心种质15个农艺性状的Shannon-w eaver和S im pson遗传多样性指数与总体资源差异不显著。表明初选核心种质能够代表全部资源的遗传多样性。     

A chromosome 21 critical region does not cause specific Down syndrome phenotypes

The "Down syndrome critical region" (DSCR) is a chromosome 21 segment purported to contain genes responsible for many features of Down syndrome (DS), including craniofacial dysmorphology. We used chromosome engineering to create mice that were trisomic or monosomic for only the mouse chromosome segment orthologous to the DSCR and assessed dysmorphologies of the craniofacial skeleton that show direct parallels with DS in mice with a larger segmental trisomy. The DSCR genes were not sufficient and were largely not necessary to produce the facial phenotype. These results refute specific predictions of the prevailing hypothesis of gene action in DS.