A late Triassic impact ejecta layer in southwestern Britain

Despite the 160 or so known terrestrial impact craters of Phanerozoic age, equivalent ejecta deposits within distal sedimentary successions are rare. We report a Triassic deposit in southwestern Britain that contains spherules and shocked quartz, characteristic of an impact ejecta layer. Inter- and intragranular potassium feldspar from the deposit yields an argon-argon age of 214 +/- 2.5 million years old. This is within the age range of several known Triassic impact craters, the two closest of which, both in age and location, are Manicouagan in northeastern Canada and Rochechouart in central France. The ejecta deposit provides an important sedimentary record of an extraterrestrial impact in the Mesozoic that will help to decipher the number and effect of impact events, the source and dynamics of the event that left this distinctive sedimentary marker, and the relation of this ejecta layer to the timing of extinctions in the fossil record.     

采用Petri网进行修正的启发式船舶电网拓扑分析

针对船舶电力系统的特点，采用一种启发式电力网络进行拓扑分析模型建立以确定电力系统网络拓扑，在此基础上借鉴电力系统故障诊断的方法,采用Petri网技术对电网进行故障定位以及对不确定信号的诊断与排除，对拓扑分析结果进行修正，以保证拓扑分析的正确性。最后以典型船舶电力系统网络为例，验证了该方法的有效性。     

Deletion in chromosome 11p associated with a hepatitis B integration site in hepatocellular carcinoma

Hepatitis B virus (HBV), a virus with known carcinogenic potential, integrates into cellular DNA during long-term persistent infection in man. Hepatocellular carcinomas isolated from viral carriers often contain clonally propagated viral DNA integrations. As small chromosomal deletions are associated with several types of carcinomas, the occurrence of chromosomal deletions in association with HBV integration in hepatocellular carcinoma was studied. HBV integration was accompanied by a deletion of at least 13.5 kilobases of cellular sequences in a human hepatocellular carcinoma. The viral DNA integration and deletion of cellular sequences occurred on the short arm of chromosome 11 at location 11p13-11p14. The cellular sequences that were deleted at the site of HBV integration were lost from the tumor cells, leaving only a single copy of the remaining cellular allele.     

Visualizing tmRNA entry into a stalled ribosome

Bacterial ribosomes stalled on defective messenger RNAs (mRNAs) are rescued by tmRNA, an approximately 300-nucleotide-long molecule that functions as both transfer RNA (tRNA) and mRNA. Translation then switches from the defective message to a short open reading frame on tmRNA that tags the defective nascent peptide chain for degradation. However, the mechanism by which tmRNA can enter and move through the ribosome is unknown. We present a cryo-electron microscopy study at approximately 13 to 15 angstroms of the entry of tmRNA into the ribosome. The structure reveals how tmRNA could move through the ribosome despite its complicated topology and also suggests roles for proteins S1 and SmpB in the function of tmRNA.     

Bacterial motility: membrane topology of the Escherichia coli MotB protein

The MotB protein of Escherichia coli is an essential component of the force generators that couple proton movement across the cytoplasmic membrane to rotation of the flagellar motors. The membrane topology of MotB was examined to explore the possibility that it might form a proton channel. MotB--alkaline phosphatase fusion proteins were constructed to identify likely periplasmic domains of the MotB molecule. Fusions distal to a putative membrane-spanning segment near the amino terminus of MotB exhibited alkaline phosphatase activity, indicating that an extensive carboxyl-terminal portion of MotB may be located on the periplasmic side of the membrane. Protease treatment of MotB in spheroplasts confirmed this view. The simple transmembrane organization of MotB is difficult to reconcile with a role as a proton conductor.     

Discovery of a predicted DNA knot substantiates a model for site-specific recombination

The mechanism of site-specific genetic recombination mediated by Tn3 resolvase has been investigated by a topological approach. Extrapolation of a detailed model of synapsis and strand exchange predicts the formation of an additional DNA product with a specific knotted structure. Two-dimensional gel electrophoresis of DNA reacted in vitro revealed a product, about 0.1 percent of the total, with the appropriate mobility. A technique for determining DNA topology by electron microscopy was improved such that less than a nanogram of DNA was required. The structure of the knot was as predicted, providing strong evidence for the model and showing the power of the topological method.     

Common pathogenetic mechanism for three tumor types in bilateral acoustic neurofibromatosis

Bilateral acoustic neurofibromatosis (BANF) is a genetic defect associated with multiple tumors of neural crest origin. Specific loss of alleles from chromosome 22 was detected with polymorphic DNA markers in two acoustic neuromas, two neurofibromas, and one meningioma from BANF patients. This indicates a common pathogenetic mechanism for all three tumor types. The two neurofibromas were among three taken from the same patient, and both showed loss of identical alleles demonstrating that the same chromosome suffered deletion in both tumors. The third neurofibroma from this patient showed no detectable loss of heterozygosity, which suggests the possibility of a more subtle mutational event that affects chromosome 22. In the two acoustic neuromas, only a portion of chromosome 22 was deleted, narrowing the possible chromosomal location of the gene that causes BANF to the region distal to the D22S9 locus in band 22q11. The identification of progressively smaller deletions on chromosome 22 in these tumor types may well provide a means to clone and characterize the defect.     

转基因玉米NK603转化体/zSSIIb内标基因二重微滴数字PCR方法的建立及应用

【目的】批准进口的转基因玉米NK603是中国转基因生物安全监管的主要对象。转基因安全监管需要标准物质和标准检测方法,建立转基因玉米NK603的数字PCR(dPCR)方法将为转基因玉米NK603的定量检测和标准物质研制提供精准测量技术。【方法】采用人工合成技术构建标准质粒分子pUC57-NK603;将NK603转化体与不同的玉米内标基因引物/探针一一组合,遴选与NK603转化体特异性PCR具有相同扩增能力的玉米内标基因PCR方法;设置二重微滴数字PCR(ddPCR)的退火温度梯度和引物/探针浓度梯度,优化二重ddPCR的反应体系和反应条件;用梯度稀释的标准质粒溶液作模板,考察二重ddPCR的检测极限、定量极限和动力学范围;将转基因玉米NK603种子粉末和非转基因玉米种子粉末混合,配制质量分数分别为100%、10%和6%的盲样,考察二重ddPCR的定量准确性。【结果】用标准质粒分子pUC57-NK603作为二重ddPCR的质控对照,通过考察二重ddPCR反应热图的微滴信号强度、阳性微滴与阴性微滴分辨率、雨滴数量、NK603转化体与内标基因拷贝数比值测量值与预期值的一致性,确定将NK603转...     

Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase

A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction. The enzyme, isolated from Thermus aquaticus, greatly simplifies the procedure and, by enabling the amplification reaction to be performed at higher temperatures, significantly improves the specificity, yield, sensitivity, and length of products that can be amplified. Single-copy genomic sequences were amplified by a factor of more than 10 million with very high specificity, and DNA segments up to 2000 base pairs were readily amplified. In addition, the method was used to amplify and detect a target DNA molecule present only once in a sample of 10(5) cells.     

A DNA segment encoding two genes very tightly linked to Huntington's disease

The discovery of D4S10, an anonymous DNA marker genetically linked to Huntington's disease (HD), introduced the capacity for limited presymptomatic diagnosis in this late-onset neurodegenerative disorder and raised the hope of cloning and characterizing the defect based on its chromosomal location. Progress on both fronts has been limited by the absence of additional DNA markers closer to the HD gene. An anonymous DNA locus, D4S43, has now been found that shows extremely tight linkage to HD. Like the disease gene, D4S43 is located in the most distal region of the chromosome 4 short arm, flanked by D4S10 and the telomere. In three extended HD kindreds, D4S43 displays no recombination with HD, placing it within 0 to 1.5 centimorgans of the genetic defect. Expansion of the D4S43 region to include 108 kilobases of cloned DNA has allowed identification of eight restriction fragment length polymorphisms and at least two independent coding segments. In the absence of crossovers, these genes must be considered candidates for the site of the HD defect, although the D4S43 restriction fragment length polymorphisms do not display linkage disequilibrium with the disease gene.     

基于SUKF与SIFT特征的红外目标跟踪算法研究

针对复杂红外背景下单一跟踪算法难以准确定位运动目标的问题,提出了基于尺度无迹卡尔曼滤波(SUKF,scale unscented Kalman filter)与尺度不变特征变换(SIFT,scale invariant featuretransform)相结合的红外运动目标跟踪方法。首先,通过SUKF算法对状态空间进行滤波估计,确定运动目标的初步位置,并以此建立局部SIFT特征检测域。其次,SIFT算法在该局部检测域内对运动目标进行特征提取与匹配,最终实现对目标的准确定位;同时,利用定位结果更新并校正SUKF的状态模型。实验结果表明,本文提出的基于SUKF-SIFT的跟踪策略与相关算法相比,体现出较好的跟踪效果与实时性能。     

Characterization and molecular cloning of a human parvovirus genome

The genome of the small human virus serologically associated with erythrocyte aplasia and erythema infectiosum (fifth disease) is shown to be a linear, nonpermuted, single-stranded DNA molecule with self-priming hairpin termini, properties which are characteristic of the genomes of the family Parvoviridae. This human parvovirus chromosome was molecularly cloned into bacterial plasmid vectors and the cloned DNA was used to explore its relatedness to other mammalian parvovirus serotypes by DNA:DNA hybridization. It is not related to the human adeno-associated viruses but does show a distant evolutionary relationship to genomes of the helper-independent parvoviruses of rodents. This strongly suggests that it is an autonomous parvovirus, and as such is the first example of a member of this group of common animal pathogens to cause disease in man.     

Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome

A random-primed complementary DNA library was constructed from plasma containing the uncharacterized non-A, non-B hepatitis (NANBH) agent and screened with serum from a patient diagnosed with NANBH. A complementary DNA clone was isolated that was shown to encode an antigen associated specifically with NANBH infections. This clone is not derived from host DNA but from an RNA molecule present in NANBH infections that consists of at least 10,000 nucleotides and that is positive-stranded with respect to the encoded NANBH antigen. These data indicate that this clone is derived from the genome of the NANBH agent and are consistent with the agent being similar to the togaviridae or flaviviridae. This molecular approach should be of great value in the isolation and characterization of other unidentified infectious agents.     

The unusual varl gene of yeast mitochondrial DNA

The var1 gene specifies the only mitochondrial ribosomal protein known to be encoded by yeast mitochondrial DNA. The gene is unusual in that its base composition is nearly 90 percent adenine plus thymine. It and its expression product show a strain-dependent variation in size of up to 7 percent; this variation does not detectably interfere with function. Furthermore, var1 is an expandable gene that participates in a novel recombinational event resembling gene conversion whereby shorter alleles are preferentially converted to longer ones. The remarkable features of var1 indicate that it may have evolved by a mechanism analogous to exon shuffling, although no introns are actually present.     

Construction of a general human chromosome jumping library, with application to cystic fibrosis

In many genetic disorders, the responsible gene and its protein product are unknown. The technique known as "reverse genetics," in which chromosomal map positions and genetically linked DNA markers are used to identify and clone such genes, is complicated by the fact that the molecular distances from the closest DNA markers to the gene itself are often too large to traverse by standard cloning techniques. To address this situation, a general human chromosome jumping library was constructed that allows the cloning of DNA sequences approximately 100 kilobases away from any starting point in genomic DNA. As an illustration of its usefulness, this library was searched for a jumping clone, starting at the met oncogene, which is a marker tightly linked to the cystic fibrosis gene that is located on human chromosome 7. Mapping of the new genomic fragment by pulsed field gel electrophoresis confirmed that it resides on chromosome 7 within 240 kilobases downstream of the met gene. The use of chromosome jumping should now be applicable to any genetic locus for which a closely linked DNA marker is available.     

Electrophoresis of flexible macromolecules: evidence for a new mode of transport in gels

Movement of macromolecules through low concentration agarose gels was investigated with linear poly(styrenesulfonate), linear DNA, star-shaped poly(styrenesulfonate), and circular DNA. Mobilities of weakly entangled flexible macromolecules were independent of molecular radius; within a homologous chemical sequence, electrophoretic separation at low field strengths depended solely on the degree of polymerization. These observations cannot be explained either by sieving or by reptation mechanisms; transport was apparently controlled by spatial variations of chain configurational entropy. Only when the chain was highly entangled did chain topology affect mobility. Evidence for entropically regulated transport clarifies how gel electrophoresis separates flexible macromolecules.     

分子标记聚合育种在作物新品种选育中的应用

分子标记的应用途径主要包括分子遗传图谱的构建和基因定位、遗传多样性与种质鉴定、分子标记辅助选择育种等3个方面。分子标记在基因聚合育种中应用的可行性已经得到证实,并在水稻白叶枯病抗性育种上取得了明显的进展,还有可能将与产量、品质或抗性等相关的不同基因或QTL聚合到一个品种之中,使品种在多个方面同时得到改良。     

A ligase-mediated gene detection technique

An assay for the presence of given DNA sequences has been developed, based on the ability of two oligonucleotides to anneal immediately adjacent to each other on a complementary target DNA molecule. The two oligonucleotides are then joined covalently by the action of a DNA ligase, provided that the nucleotides at the junction are correctly base-paired. Thus single nucleotide substitutions can be distinguished. This strategy permits the rapid and standardized identification of single-copy gene sequences in genomic DNA.