Modification of a viral satellite DNA-based gene silencing vector and its application to leaf or flower color change in Petunia hybrida

Gene silencing conserved in plants and animals is mediated by mechanisms that involve sequence- specific RNA degradation[1,2]. Gene silencing has been proven to play an important role in the study of gene function. Recently, a procedure known as virus ind…     

DNA分子标记在真菌系统分类与鉴定中的应用

近年来分子生物学技术的发展,为从核酸分子水平上研究真菌的遗传本质,为真菌的进化研究、系统分类和鉴定开辟了新的途径.概述了几种主要的DNA分子标记的基本原理和特点,以及在真菌鉴定与分类中的应用.     

一种PCR快速鉴定重组体DNA阳性克隆及插入方向的方法

当外源DNA以非定向的方式插入克隆载体时,需用测序或酶切的方法确定插入方向,上述两种方法或昂贵或费力。描述了一种快速、简便的PCR法,即利用载体的通用引物和目的片段的两个特异性引物直接扩增重组菌落,从而达到鉴别DNA插入方向的目的。     

农杆菌介导的胆碱脱氢酶基因(betA)转化小麦及影响转化效率的因素

用LBA4404／pCDH,Agl 1／pUNN2和Ag;P(无质粒)3种菌株分别转化小麦品种济南177、99P、核生3号幼胚诱导的愈伤组织以及济南177的幼胚．其中以胚性愈伤组织为外植体,获得了转基因植株．PCR和PCR-Southem分析证实转化植株中包含了外源基因．通过染色体分析,发现胚性愈伤组织在农杆菌LBA4404侵染后形成的染色体削减比经Agl1侵染和未经感染的对照形成的染色体削减程度更大,甚至发现较多的染色体断片．分析了农杆菌菌株,材料的基因型,外植体类型,培养时间和温度以及筛选的时间和周期对于转化效率的影响．     

不同破壁方法对活性污泥总DNA提取效果的影响

分别采用酶法、化学法、珠磨匀浆法和反复冻融法对活性污泥样品进行破壁,提取总DNA,并通过提取的核酸总量、纯度、片段分布情况、是否含有聚合酶链反应抑制剂以及基因间隔序列(ITS)扩增产物多态性等5个指标来评价不同的细胞破壁方法对活性污泥总DNA提取效果的影响。结果表明,化学法的优势最明显,提取的总量最多,大片段提取效果好,虽然杂质较多,但不影响聚合酶链反应,方法的偏倚性最小。     

文蛤(Meretrix meretrix)地理种群ISSR分子标记的初步研究

利用ISSR(InterSimpleSequenceRepeate)技术对文蛤(Meretrixmeretrix)江苏和辽宁两个地理种群进行了PCR扩增.从100条ISSR引物中筛选出引物13条,每条引物检测出位点数1到8.平均每条引物可检测到位点数4 6个.实验结果表明:江苏文蛤的位点多态性(80 7%)高于辽宁文蛤的位点多态性(68 4%);种群内平均遗传距离分别为0 3105±0 090和0 2658±0 044,江苏文蛤也高于辽宁文蛤.从筛选得到的引物可以发现,文蛤简单重复序列中A、G碱基含量较高.通过对不同实验条件的对比,对ISSR PCR反应体系进行了优化.     

DNA synthesis in the nuclei of differentiating muscle fibers of the silkworm,Bombyx mori L.

Summary Incorporation of³H-thymidine (³HTdr) into the nuclei of myofibril-containing myofibers of larvae of the silkworm,Bombyx mori, was shown by means of light microscope (LM) and electron-microscope (EM) autoradiography. The number of DNA-synthesizing myonuclei attains 42% 12–18 h after each molt. Thus in the developing silkworm DNA replication and myofibrillogenesis are coexisting and not mutually exclusive processes as is the rule in vertebrate somatic myogenesis.     

DNA in wheat seeds from European archaeological sites

We have used hybridization analysis to detect ancient DNA in wheat seeds collected from three archaeological sites in Europe and the Middle East. One of these samples, carbonizedT. spelta dated to the first millennium BC, has yielded PCR products after amplification with primers directed at the leader regions of the HMW (high molecular weight) glutenin alleles. Sequences obtained from these products suggest that the DNA present in the Danebury seeds is chemically damaged, as expected for ancient DNA, and also indicate that it should be possible to study the genetic variability of archaeological wheat by ancient DNA analysis. Finally, we describe a PCR-based system that enables tetraploid and hexaploid wheats to be distinguished.     

寡聚酰胺类DNA识别分子研究进展

从有机化学的角度综述了近年来寡聚酰胺类DNA识别分子的设计、合成、识别及应用等方面的研究进展情况．     

Genetic variation in the New World: Ancient teeth,bone, and tissue as sources of DNA

Examination of ancient and contemporary Native American mtDNA variation via diagnostic restriction sites and the 9-pb Region V deletion suggests a single wave of migration into the New World. This is in contrast to data from Torroni et al.³⁴ which suggested two waves of migration into the New World (the NaDene and Amerind). All four founding lineage types are present in populations in North, Central, and South America suggesting that all four lineages came over together and spead throughout the New World. Ancient Native American DNA shows that all four lineages were present before European contact in North America, and at least two were present in South America. The presence of all four lineages in the NaDene and the Amerinds argues against separate migrations founding these two groups, although admixture between the groups is still a viable explanation for the presence of all four types in the NaDene.