减数分裂型黏连蛋白亚基SMC1β和STAG3在染色体运动中的功能启动时期研究

维持姐妹染色单体的黏着是保证染色体正确附着在纺锤体上,从而实现正确分离的关键。黏连蛋白在有丝分裂和减数分裂过程中对染色体结构和染色体分离具有重要作用。在迄今为止发现的四种减数分裂型黏连蛋白亚基RAD 21L、REC8、SMC1β和STAG3中,RAD 21L是具有配对同源染色体的主导蛋白质,在减数分裂前细线期同源染色体配对的启动过程中起着中心作用。为揭示SMC1β和STAG3在减数分裂期染色体运动中的作用起始点,观察了异位表达蛋白在体细胞中对染色体运动的影响。结果显示:与对照(未转染)细胞相比,异位表达的GFP标记SMC1β或STAG3对两个FISH荧光信号之间的距离没有显著影响,这两个FISH荧光信号代表核内一对X染色体或一对11染色体。研究结果表明:至少在减数分裂期同源染色体配对过程开始之前,SMC1β或STAG3都不履行各自的功能。     

节节麦和黑麦杂种胚再生植株的分子细胞遗传学分析

利用杂种幼胚愈伤组织诱导和再生技术产生了节节麦和黑麦杂种幼胚无性系.胚拯救直接成苗植株、继代培养60d的再生植株和继代培养420d的再生杂种植株PMCs减数分裂MI染色体构型分别为10.82I+1.57II+0.01III,9.96I+1.59II+0.25III+0.03IV,7.59Ⅰ+3.10Ⅱ+0.05Ⅲ+0.01IV.继代培养420d左右胚再生无性系其减数分裂MI染色体配对频率大幅度提高,利用原位杂交技术分析表明其染色体配对频率提高和染色体结构和数量变异无关.提示体细胞无性系变异可能是增加部分同源染色体配对,增加属间种基因交流的一个新途径.     

小麦染色体配对基因Ph1结构剖析及调控机制研究进展

近年来,关于小麦抑制部分同源染色体配对基因Ph1的研究有了突破性进展.本文对该基因的结构和调控机理的最新研究进行综述.通过创造和分子标记鉴定Ph1缺失突变体,利用分子生物学及比较基因组学技术,该基因位点被界定于5BL上一个2.5 Mb的区域内,含有一个类cdk基因簇,且在该类cdk基因簇中插入一个亚端粒异染色质片段.细胞学研究显示,Ph1基因通过控制亚端粒的互作启始染色体识别和配对伙伴选择.与此同时,生物信息学揭示,这些类cdk基因与人类和老鼠的cdk2基因高度同源,它们与细胞周期中DNA复制、染色质凝集、碱基错配修复等事件相关.减数分裂时,该基因位点通过"感知"染色体的同源性程度而触发染色质的构象变化,从而控制染色体的配对和重组.此外,小麦中可能存在一种与Ph1相关的类似于酵母中的粗线期检查点机制.预测未来的研究将可能集中在Ph1对染色体同源性的"感知"机制、Ph1的开启与关闭、植物减数分裂重组的忠实性及减数分裂过程的检查点机制等方面.     

利用小麦5B效应引入外源基因的研究（Ⅰ）易位系的培育

ｄｕｒｕｍ小麦的代换系ｄｉ－ｓｕｂ５Ｄ（５Ｂ）与添加系ｄｉ－ａｄｄｅ４ｔｓ杂交，再用ｄｉ－ｓｕｂ５Ｄ（５Ｂ）进行回交，在自交后代中选育出了易位系１０３２。该易位系染色体数２ｎ＝２８，表现型为非蜡质。这一结果证明了在ｄｕｒｕｍ小麦中也可以利用５Ｂ染色体效应，通过诱发部份同源染色体间的配对，获得易位体。     

染色体的空间分布

本文主要就细胞分裂周期中的同一个染色体组内染色体的空间分布,二倍体或导源多倍体中同源染色体组间的空间分布,同源多倍体中同源染色体组间的空间分布,异源多倍体中部分同源染色体组间的空间分布以及这些分布的机制进行述评,并对研究这个问题的方法和意义进行讨论。     

黄果西番莲染色体G—显带和G—带带型

用正红花油（ＲＦＯ）去壁，Ｇｉｅｍｓａ染色法对黄果西番莲的染色体进行了Ｇ－显带研究和Ｇ－带带型分析。结果表明，全部９对染色体（２ｎ＝１８）均显示出清晰的丰富的Ｇ－带带纹，但各条染色体的带纹数目随着有丝分裂时期的推进而逐渐减少，前期带纹数量多，早中期次之，中期最少或无带；各对同源染色体的两个成员之间带纹的位置、数目、大小、着色深浅和带间宽窄，彼此基本相似，可较准确配对，而非同源染色体之间的带型特征则     

减数分裂是三个遗传基本规律的细胞学基础

<正> 减数分裂是生殖细胞的成熟分裂,在减数分裂过程中:一方面完成了染色体数目减半(在整个减数分裂过程中,染色体复制一次而细胞连续分裂两次);另一方面同源染色体的特殊行为:同源染色体的联会和分离;同源染色体的非姐妹染色单体之间的交叉互换;非同源染色体之间的自由组合,是三个遗传基本规律的细胞学基础。     

AGP转基因粳稻产量性状在低世代中的遗传与变异

云南粳稻合系３８、合系４２经ＡＧＰ基因导入后,株高,有效穗数,实粒数,千粒重等产量性状的群体变异频率在Ｔ１和Ｔ２代中显著增高,由于水稻自花授粉的特性,导入目的基因的两条同源染色体配对的频率也较高,因此在代世代中已经出现了综合性状较好的株系。     

异细胞质在八倍体小黑麦育种上利用的研究

利用１５个异细胞质中国春及对照中国春分别与黑麦杂交，结实率有所不同，出苗率差异显著，异细胞质对Ｆ１减数分裂中期Ⅰ梁色体Ａｅ．ｓｈａｒｏｎｅｓｉｅ和Ａｅ．ｂｖｉｃｏｒｎｉｓ细胞分别促进和抑制中国春与黑麦Ｆ１部分同源染色体配对。     

小麦-多枝赖草单体异附加材料减数分裂期外源染色体的行为研究

通过细胞学观察和基因组原位杂交技术分析了小麦-多枝赖草单体异附加材料减数分裂期外源染色体的遗传行为.研究结果表明:在减数分裂中期,附加的多枝赖草染色体在一定程度上干扰小麦同源染色体的正常配对,出现了未配对的小麦染色体;在减数分裂后期,30.2%～35.6%的外源染色体随机走向一极,19.5%～20.9%的外源染色体提早分裂,多枝赖草染色体落后和断裂的频率分别为19.8%～21.1%和7.9%～10.5%,同时有少量小麦染色体也发生了不规则分裂现象,表明附加的多枝赖草染色体也影响了小麦染色体后期的正常分离,这为小麦与多枝赖草染色体间产生易位提供了条件.     

褐家鼠染色体减数分裂联会复合体的研究

以界面铺张－硝酸银和磷钨酸（ＰＴＡ）染色技术，对褐家鼠（Ｒａｔｔｕｓｎｏｒｖｅｇｉｃｕｓ）减数分裂前期精母细胞联会复合体（ＳＣ）进行了显微和亚显微结构的观察．银染和ＰＴＡ染色都清楚地显示出褐家鼠ＳＣ的结构和减数分裂的行为，但ＰＴＡ染色的ＳＣ标本，着丝位区域较为明显．ＳＣｓ核型与有丝分裂染色体核型有较好的一致性．     

犏牛精母细胞联会复合体与雄性不育关系的研究

采用界面铺展硝酸银染色技术，研究了公犏牛的精母细胞联合复合体（ｓｙｎａｐｔｏｎｅｍａｌｃｏｍｐｌｅｘ，ＳＣ），结果表明：（１）在减数分裂过程中仅有少数精母的常染色体能形成ＳＣ结构，Ｘ、Ｙ染色体轴 围虽积聚有许多微小的深色颗粒状物质，但未见Ｘ、Ｙ染色体的明显联会（２）在常染色体的联会中，有较多的三价体、插入环等联会异常现象；（３）常染色体ＳＣ相对长度与体细胞染色体的相对长度间有显著相关性（Ｒ＝０９６     

Single-molecule imaging of DNA pairing by RecA reveals a three-dimensional homology search

DNA breaks can be repaired with high fidelity by homologous recombination. A ubiquitous protein that is essential for this DNA template-directed repair is RecA. After resection of broken DNA to produce single-stranded DNA (ssDNA), RecA assembles on this ssDNA into a filament with the unique capacity to search and find DNA sequences in double-stranded DNA (dsDNA) that are homologous to the ssDNA. This homology search is vital to recombinational DNA repair, and results in homologous pairing and exchange of DNA strands. Homologous pairing involves DNA sequence-specific target location by the RecA-ssDNA complex. Despite decades of study, the mechanism of this enigmatic search process remains unknown. RecA is a DNA-dependent ATPase, but ATP hydrolysis is not required for DNA pairing and strand exchange, eliminating active search processes. Using dual optical trapping to manipulate DNA, and single-molecule fluorescence microscopy to image DNA pairing, we demonstrate that both the three-dimensional conformational state of the dsDNA target and the length of the homologous RecA-ssDNA filament have important roles in the homology search. We discovered that as the end-to-end distance of the target dsDNA molecule is increased, constraining the available three-dimensional (3D) conformations of the molecule, the rate of homologous pairing decreases. Conversely, when the length of the ssDNA in the nucleoprotein filament is increased, homology is found faster. We propose a model for the DNA homology search process termed 'intersegmental contact sampling', in which the intrinsic multivalent nature of the RecA nucleoprotein filament is used to search DNA sequence space within 3D domains of DNA, exploiting multiple weak contacts to rapidly search for homology. Our findings highlight the importance of the 3D conformational dynamics of DNA, reveal a previously unknown facet of the homology search, and provide insight into the mechanism of DNA target location by this member of a universal family of proteins.     

The Ph1 locus is needed to ensure specific somatic and meiotic centromere association

The correct pairing and segregation of chromosomes during meiosis is essential for genetic stability and subsequent fertility. This is more difficult to achieve in polyploid species, such as wheat, because they possess more than one diploid set of similar chromosomes. In wheat, the Ph1 locus ensures correct homologue pairing and recombination. Although clustering of telomeres into a bouquet early in meiosis has been suggested to facilitate homologue pairing, centromeres associate in pairs in polyploid cereals early during floral development. We can now extend this observation to root development. Here we show that the Ph1 locus acts both meiotically and somatically by reducing non-homologous centromere associations. This has the effect of promoting true homologous association when centromeres are induced to associate. In fact, non-homologously associated centromeres separate at the beginning of meiosis in the presence, but not the absence, of Ph1. This permits the correction of homologue association during the telomere-bouquet stage in meiosis. We conclude that the Ph1 locus is not responsible for the induction of centromere association, but rather for its specificity.     

哺乳动物克隆的现状和研究进展

 哺乳动物细胞克隆是20世纪末生命科学领域最引人注目的高新技术,该技术对于优良种畜的复制、减少试验用动物数目、动物遗传多样性保存及濒危动物挽救、转基因动物培育等方面具有重要意义。近年来克隆技术发展迅速,多种哺乳动物相继克隆成功,但也存在克隆效率太低、克隆动物表型正常而实质异常的问题。本文详细阐述了克隆效率太低、克隆动物表型正常而实质异常问题,介绍了当前动物克隆技术的发展现状,并对动物克隆涉及的技术进行了总结和概括,着重介绍了卵母细胞的去核方法和重组胚的构建方法。     

抗病毒中药复方制剂对雏鸡免疫功能影响的研究

探讨抗病毒中药对雏鸡免疫功能的影响,为药理作用和临床合理用药提供科学依据.采用水萃取法制备抗病毒中药复方浸膏制剂,将实验动物随机分为阴性对照组、阳性对照组、低剂量中药组、中剂量中药组和高剂量中药组,在免疫接种NDV-Lasota株灭活疫苗的基础之上,以NDV-Ab(鸡新城疫病毒抗体)、CD4+/CD8+T淋巴细胞比值为检测指标,研究中药复方制剂对雏鸡免疫功能的影响.实验结果表明:高剂量中药组在连续给药5d后,能够促进实验雏鸡在疫苗接种后7-14d的NDV-Ab水平急剧上升,14d达到最高(27.8073ng/L),极显著高于阳性对照组的17.8183ng/L(P0.01);3个剂量中药组均能提高实验雏鸡在疫苗接种后7-14d外周血T淋巴细胞CD4+/CD8+比值的作用,14d达到最高,高于阳性对照组(P0.05).结论:抗病毒中药复方制剂具有诱导实验雏鸡体液免疫应答的作用,诱导作用可能与中药剂量及机体的免疫状态有关.     

Formation and resolution of recombination intermediates by E. coli RecA and RuvC proteins.

The recombination of DNA molecules has been reconstituted in vitro using two purified enzymes from Escherichia coli. RecA protein catalyses homologous pairing and strand exchange reactions to form intermediate DNA structures that are acted upon by RuvC. The newly identified RuvC protein resolves the intermediates by specific endonucleolytic cleavage to produce recombinant DNA molecules.     

Germline stem cells and follicular renewal in the postnatal mammalian ovary

A basic doctrine of reproductive biology is that most mammalian females lose the capacity for germ-cell renewal during fetal life, such that a fixed reserve of germ cells (oocytes) enclosed within follicles is endowed at birth. Here we show that juvenile and adult mouse ovaries possess mitotically active germ cells that, based on rates of oocyte degeneration (atresia) and clearance, are needed to continuously replenish the follicle pool. Consistent with this, treatment of prepubertal female mice with the mitotic germ-cell toxicant busulphan eliminates the primordial follicle reserve by early adulthood without inducing atresia. Furthermore, we demonstrate cells expressing the meiotic entry marker synaptonemal complex protein 3 in juvenile and adult mouse ovaries. Wild-type ovaries grafted into transgenic female mice with ubiquitous expression of green fluorescent protein (GFP) become infiltrated with GFP-positive germ cells that form follicles. Collectively, these data establish the existence of proliferative germ cells that sustain oocyte and follicle production in the postnatal mammalian ovary.     

Programmed cell death in developing human fetal CNS

The spatial and temporal distributions of programmed cell death (PCD) in developing central nervous system (CNS) of human fetuses ranging from 12 to 39 weeks of gestation were investigated using techniques of flow cytometry and terminal transferase-mediated nick end labeling (TUNEL). The results showed that PCD did occur in every representative brain region of all fetuses examined in different stages. It was found that there were two peaks of PCD appearing at the 12th and 39th weeks respectively, which suggested that the first peak of apoptosis may be involved in the selective elimination of neurons overproduced during the early development and the second may play an important role in establishing the correct neuronal circuitry.