耐辐射奇球菌抗辐射机制的研究进展

耐辐射奇球菌是迄今为止发现的对辐射抗性最强的原核生物,是研究DNA损伤与修复的模式生物。根据国内外对耐辐射奇球菌研究最新成果,从该细菌的生理生化特征和遗传学特性、诱变所致的损伤与其高效的修复机制,及应用方面的研究进行了综述,探讨了其研究价值和应用前景。     

Ringlike structure of the Deinococcus radiodurans genome: a key to radioresistance?

The bacterium Deinococcus radiodurans survives ionizing irradiation and other DNA-damaging assaults at doses that are lethal to all other organisms. How D. radiodurans accurately reconstructs its genome from hundreds of radiation-generated fragments in the absence of an intact template is unknown. Here we show that the D. radiodurans genome assumes an unusual toroidal morphology that may contribute to its radioresistance. We propose that, because of restricted diffusion within the tightly packed and laterally ordered DNA toroids, radiation-generated free DNA ends are held together, which may facilitate template-independent yet error-free joining of DNA breaks.     

耐辐射菌中抗辐射相关基因或酶的研究进展

耐辐射菌对电离辐射、紫外线和一些DNA损伤剂等具有极强的抵抗能力,相关研究表明,耐辐射菌超强的抗辐射作用机制主要存在3个方面,即DNA损伤的高效修复、抗氧化作用以及特殊的生存方式。该研究就耐辐射菌的抗辐射作用相关基因或酶进行了简要综述,并对其应用前景进行了展望。     

耐辐射球菌DNA修复机制研究新进展

耐辐射球菌是迄今为止发现的最耐辐射的原核生物,是研究DNA损伤与修复的模式生物.根据国内外实验室和本实验在耐辐射球菌研究上取得的最新研究成果,本文从该细菌的结构特征、分子防御机制、重要修复基因、基因组学和蛋白质组学等方面综述了耐辐射球菌在DNA修复机制方面取得的进展,探讨了未来揭示该细菌独特高效的DNA修复分子机理可能采取的途径.     

耐辐射异常球菌DNA损伤修复重要功能蛋白的研究进展

耐辐射异常球菌(DR)具有对致死剂量的电离辐射极端的抗性,但对其超强辐射抗性的分子机制仍缺乏深入了解。耐辐射异常球菌基因组分析显示其拥有许多与修复相关的基因或蛋白,转录组学和蛋白组学分析等研究表明该菌能通过复杂的代谢途径控制的网络系统有效地调整并修复DNA。近年来已分析和鉴定了许多与DNA损伤修复,特别是辐射诱导的DNA双链断裂修复相关的蛋白,这些功能蛋白的研究有助于我们加深对其极端抗性机理的认识。主要针对DR双链断裂修复系统中重要功能蛋白的研究进展进行了概述,以期为DNA修复机理的基础研究及应用研究奠定基础。     

Bacteriophytochromes: phytochrome-like photoreceptors from nonphotosynthetic eubacteria

Phytochromes are a family of photoreceptors used by green plants to entrain their development to the light environment. The distribution of these chromoproteins has been expanded beyond photoautotrophs with the discovery of phytochrome-like proteins in the nonphotosynthetic eubacteria Deinococcus radiodurans and Pseudomonas aeruginosa. Like plant phytochromes, the D. radiodurans receptor covalently binds linear tetrapyrroles autocatalytically to generate a photochromic holoprotein. However, the attachment site is distinct, using a histidine to potentially form a Schiff base linkage. Sequence homology and mutational analysis suggest that D. radiodurans bacteriophytochrome functions as a light-regulated histidine kinase, which helps protect the bacterium from visible light.     

耐辐射球菌抑菌圈试验过程中出现的问题与分析

[目的]分析耐辐射球菌抑菌圈试验过程中出现的问题并提出解决方法。[方法]采用已发表的基因组序列作参考,对来自于耐辐射球菌的所有菌株均用TCY培养基或者TGY平板在30℃条件下进行培养,参照Debabrota等的方法进行抑菌茵圈试验,所用突变体源于以前的试验。[结果]分析认为,在做耐辐射球菌抑菌圈试验的过程中,首先需要注意的是严格灭菌,每次重新拿到超净工作台上的离心管和钢圈都要进行严格的灭菌处理。试验前对上清液要进行高速离心,以防止在上清液中混有菌体,从而造成结果不准确。最好在进行抑菌圈试验以前,先对上清液中是否含有菌体做一个前期试验,以免造成后面不必要的麻烦。[结论]该研究所提出的解决方法对做好以后的试验具有借鉴意义.     

耐辐射异常球菌黄嘌呤脱氢酶在氧化胁迫反应中的作用

黄嘌呤脱氢酶广泛分布在真核生物、细菌和古细菌中,是催化氧化嘌呤、蝶啶和醛类等多种杂环分子的氧化还原酶类,与氮同化、激素代谢、衰老的调控及活性氧产生等过程相关.耐辐射异常球菌(Deinococcus radiodurans R1)具有超强的氧化胁迫抗性和氧保护机制,其基因组含有完整的黄嘌呤脱氢酶编码基因xdhABC.为探...     

耐辐射球菌离子注入抗性的初步研究

[目的]为深入分析耐辐射球菌的抗性机制奠定基础。[方法]以大肠杆菌为对照,研究经γ射线辐射和N+注入后耐辐射球菌的剂量-存活率曲线和剂量-生物量曲线,并分析总蛋白的变化情况。[结果]注入N+后,耐辐射球菌的存活率和生物量均呈降-升-降的马鞍型曲线。耐辐射球菌在各个辐射剂量下的存活率高于大肠杆菌,其生物量总体上也多于大肠杆菌,说明耐辐射球菌具有较高抗性。在低剂量N+注入后,总蛋白的含量变化不明显,在高剂量时蛋白含量增加明显。[结论]耐辐射球菌在高剂量下具有一定的修复能力。     

Recognition of thymine adenine.base pairs by guanine in a pyrimidine triple helix motif

Oligonucleotide recognition offers a powerful chemical approach for the sequence-specific binding of double-helical DNA. In the pyrimidine-Hoogsteen model, a binding size of greater than 15 homopurine base pairs affords greater than 30 discrete sequence-specific hydrogen bonds to duplex DNA. Because pyrimidine oligonucleotides limit triple helix formation to homopurine tracts, it is desirable to determine whether oligonucleotides can be used to bind all four base pairs of DNA. A general solution would allow targeting of oligonucleotides (or their analogs) to any given sequence in the human genome. A study of 20 base triplets reveals that the triple helix can be extended from homopurine to mixed sequences. Guanine contained within a pyrimidine oligonucleotide specifically recognizes thymine.adenine base pairs in duplex DNA. Such specificity allows binding at mixed sites in DNA from simian virus 40 and human immunodeficiency virus.     

耐辐射异常球菌K+吸收蛋白ktrA基因功能研究

耐辐射异常球菌(D. radiodurans R1)ktrA基因(DR_1666)编码的K+吸收蛋白在盐胁迫和渗透胁迫中起着重要作用。采用QRT\|PCR分析表明在盐和D\|山梨糖醇胁迫条件下,野生型菌株中ktrA基因表达均显著上调。同时采用融合PCR方法和同源重组技术,构建ktrA基因缺失突变株(ΔktrA),以野生型菌株DR做对照,分别用不同浓度的氯化钾和D\|山梨糖醇对突变株进行胁迫处理。结果显示,突变株ΔktrA对氯化钾和D\|山梨糖醇的敏感性明显高于野生型,表明ktrA基因在菌株耐受高盐和高渗透胁迫时起重要作用。该结果为深入研究D. radiodurans R1菌株中ktrA基因的功能提供了很好的基础。     

Accumulation of Mn(II) in Deinococcus radiodurans facilitates gamma-radiation resistance

Deinococcus radiodurans is extremely resistant to ionizing radiation. How this bacterium can grow under chronic gamma radiation [50 grays (Gy) per hour] or recover from acute doses greater than 10 kGy is unknown. We show that D. radiodurans accumulates very high intracellular manganese and low iron levels compared with radiation-sensitive bacteria and that resistance exhibits a concentration-dependent response to manganous chloride [Mn(II)]. Among the most radiation-resistant bacterial groups reported, Deinococcus, Enterococcus, Lactobacillus, and cyanobacteria accumulate Mn(II). In contrast, Shewanella oneidensis and Pseudomonas putida have high iron but low intracellular manganese concentrations and are very sensitive. We propose that Mn(II) accumulation facilitates recovery from radiation injury.     

六倍体裸燕麦(Avena nuda L.)染色体组型分析

本实验对六倍体裸燕素(Avena nuda L.)进行染色体组型研究。该品种染色体数目为:2n=6x=42,是稳定的六倍体。并对其染色体形态特征进行了研究。六倍体裸燕麦(代1109)的21对染色体中,有10对为中着丝点染色体(M),有8对近中着丝点染色体(SM),有3对具有随体的染色体(SAt)。其染色体组型为:2n=20M+16SM+6SAt。为燕麦分类提供了细胞学依据。     

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.     

耐辐射球菌PPrI突变株的蛋白质组学分析

PprI由dr0167编码,在耐辐射球菌的极端抗性中起重要作用.pprI突变株对于电离辐射、紫外线辐射、丝裂霉素C等都很敏感.与正常培养的野生型菌株比较,正常培养的pprI突变株的双向图谱有明显的变化,其中有18个蛋白质表达量增加,7个蛋白质表达量减少.在有差异的25个蛋白质中,我们用MALDI-TOF质谱仪成功鉴定了13个,这些蛋白质在细胞中行使不同的功能,包括:①糖类转运与代谢;②能量的产生与转化;③无机离子转运与代谢;④氨基酸转运与代谢;⑤翻译后修饰,蛋白代谢,分子伴侣及一些未知的功能等.研究表明,PprI不仅调控与DNA修复直接相关的基因表达,而且调控一个极其复杂的网络系统,这个系统包括行使各种生物学的功能蛋白.     

Molecular analysis of a constitutional X-autosome translocation in a female with muscular dystrophy

The gene responsible for Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) maps to the X chromosome short arm, band Xp21. In a few females with DMD or BMD, the Xp21 region is disrupted by an X-autosome translocation. Accumulating evidence suggests that the exchange has physically disrupted the DMD/BMD locus to cause the disease. One affected female with a t(X;21)(p21;p12) translocation was studied in detail. The exchange points from both translocation chromosomes were cloned, restriction-mapped, and sequenced. The translocation is reciprocal, but not conservative. A small amount of DNA is missing from the translocated chromosomes; 71 to 72 base pairs from the X chromosome and 16 to 23 base pairs from the 28S ribosomal gene on chromosome 21.     

Metagenomics to paleogenomics: large-scale sequencing of mammoth DNA

We sequenced 28 million base pairs of DNA in a metagenomics approach, using a woolly mammoth (Mammuthus primigenius) sample from Siberia. As a result of exceptional sample preservation and the use of a recently developed emulsion polymerase chain reaction and pyrosequencing technique, 13 million base pairs (45.4%) of the sequencing reads were identified as mammoth DNA. Sequence identity between our data and African elephant (Loxodonta africana) was 98.55%, consistent with a paleontologically based divergence date of 5 to 6 million years. The sample includes a surprisingly small diversity of environmental DNAs. The high percentage of endogenous DNA recoverable from this single mammoth would allow for completion of its genome, unleashing the field of paleogenomics.     

Chromosome localization of highly repetitive human DNA's and amplified ribosomal DNA with restriction enzymes

Restriction endonucleases cut and partially removed DNA throughout fixed air-dried human metaphase chromosomes. Some enzymes produced a G-banding pattern; some revealed the presence of multiple chromosome-specific classes of highly repetitive DNA in C-band heterochromatin. Enzymes that produced the informative C-band patterns had recognition sequences that were four or five, but not six, base pairs long and did not contain a cytosine-guanine doublet. In both rat and human chromosomes, regions containing amplified ribosomal RNA genes were specifically removed by the restriction endonuclease Msp I.     

野燕麦的C带核型研究

用Ｃ－分带技术鉴定六倍体燕麦的２１对染色体并建立燕麦的Ｃ带核型模式,发现燕麦的２１对染色体分成３组,每组７对,其中Ｃ组染色体分布有较多的异染色质区域,染色较深,很容易与Ａ组和Ｄ组区分开。Ａ组和Ｄ组染色体染色较浅,尽管每对染色体都有自己的特征带,但要将１４对浅染的染色体归入不同的部分同源染色体组还需要积累更多的细胞学和分子学的资料