Telomeric repeat from T. thermophila cross hybridizes with human telomeres

The ends (telomeres) of eukaryotic chromosomes must have special features to ensure their stability and complete replication. Studies in yeast, protozoa, slime moulds and flagellates show that telomeres are tandem repeats of simple sequences that have a G-rich and a C-rich strand. Mammalian telomeres have yet to be isolated and characterized, although a DNA fragment within 20 kilobases of the telomeres of the short arms of the human sex chromosomes has been isolated. Recently we showed that a chromosome from the fission yeast Schizosaccharomyces pombe could, in some cases, replicate as an autonomous mini-chromosome in mouse cells. By extrapolation from other systems, we reasoned that mouse telomeres could be added to the S. pombe chromosome ends in the mouse cells. On setting out to test this hypothesis we found to our surprise that the telomeric probe used (containing both the S. pombe and Tetrahymena thermophila repeats) hybridized to a series of discrete fragments in normal mouse DNA and DNA from a wide range of eukaryotes. We show here that the sequences hybridizing to this probe are located at the telomeres of most, if not all, human chromosomes and are similar to the Tetrahymena telomeric-repeat component of the probe.     

“Chi-Fang-Qi”: Is it the aureole of an ancient supernova?

An annotation for the statement about JingXing (Great Star) in the most famous Chinese ancient historical book Ski Ji Historical Records authored by Sima Qian reads as follows: “There was a red aureole (“Chi-Fang-Qi”) which connected with a deep-color aureole (“Qing-Fang-Qi”). Two yellaw stars were in the red aureole,one yellow star was in the deep-color aureole”. The author of the annotation was Meng Kang, an officer as well as a scholar of the Wei State in the Three Kingdoms period. The same records are to be found in two other Chinese histurical books compiled by the authors of later ages in Tang Dynasty——Jin History and Sui History. The scene illustrated by the record of “Chi-Fang-Qi” is very similar to the pictures of SN1987A taken by the Hubble Space Telescope. Let it be noted that although the ancient authors were often with serious wrong understandings of some astronomical phenomena such as supernovae, their depictions about natural phenomena they witnessed were mostly objective, and it was unlikely that they fictionally made the record of “Chi-Fang-Qi”. We infer that the ancient observers might have ever watched a very bright supernnva with an aureole structure in its remnant and then left such a record. Meng Kang perhaps was the first to give the record about“Chi-Fang-Qi”, and he might have lived in the period from about AD 180 to AD250, very near to AD 185. so, here the supernova the ancient people watched was most likely the one in the year of AD 185. This supposition is consistent with the distance parameter of the most possible remnant of SN185 derived by some modern measurements. If the correlation between the record of “Chi-Fang-Qi” and the supernova remnant can he further proven true, it would be an important verification of the modern theory about stellar evolution.     

ISS： Efficient Search Scheme Based on Immune Method in Modern Unstructured Peer-to-Peer Networks

Flooding is the most famous technique for locating contents in unstructured P2P networks. Recently traditional flooding has been replaced by more efficient dynamic query （DQ） and different variants of such algorithms. Dynamic query is a new flooding technique which could estimate a proper time-to-live （TTL） value for a query flooding by estimating the popularity of the searched files, and retrieve sufficient results under controlled flooding range for reducing network traffic. However, all DQ-like search algorithms are ＂blind＂ so that a large amount of redundant messages are caused. In this paper, we proposed a new search scheme, called Immune Search Scheme （ISS）, to cope with this problem. In ISS, an immune systems inspired concept of similarity-governed clone proliferation and mutation for query message movement is applied. Some assistant strategies, that is, shortcuts creation and peer traveling are incorporated into ISS to develop ＂immune memory＂ for improving search performance, which can make ISS not be blind but heuristic.     

Cloning of human telomeres by complementation in yeast

Telomeres confer stability on chromosomes by protecting them from degradation and recombination and by allowing complete replication of the end. They are genetically important as they define the ends of the linkage map. Telomeres of lower eukaryotes contain short repeats consisting of a G-rich and a C-rich strand, the G-rich strand running 5'-3' towards the telomere and extending at the end. Telomeres of human chromosomes share characteristics with those of lower eukaryotes including sequence similarity as detected by cross-hybridization. Telomeric repeats from many organisms can provide telomere function in yeast. Here we describe a modified yeast artificial chromosome (YAC) vector with only one telomere which we used to clone human telomeres by complementation in yeast. YACs containing human telomeres were identified by hydridization to an oligonucleotide of the trypanosome telomeric repeat. A subcloned human fragment from one such YAC is immediately subtelomeric on at least one human chromosome.     

Molecular cloning of human telomeres in yeast

Telomeres are the DNA sequences found at the ends of linear chromosomes. They define the boundaries of the genetical and physical maps of such chromosomes and so are particularly important for the complete mapping of large genomes that is now being attempted. Telomeres have been intensively studied in the yeast Saccharomyces cerevisiae and in ciliated protozoa: in these organisms the telomeric DNA consists of arrays of tandemly repeated short sequences in which one strand is guanosine-rich and oriented 5' to 3' towards the chromosome end. The conservation of these structural features is reflected in the observation that telomeric DNA from a variety of protozoa will function as telomeres on artificial linear mini-chromosomes in yeast. Tandem arrays of the sequence TTAGGG have been identified at the telomeres of humans and other mammals and also of trypanosomes. This indicates that the structural features of telomeres are conserved between higher and lower eukaryotes and implies that human telomeric DNA could function in yeast. I have used this idea to develop a strategy to isolate a specific human telomere as a molecular clone in yeast and have devised a simple and effective way of cloning other human telomeres and their associated sequences.     

Identification of an AFLP marker linked to the stripe rust resistance geneYr10 in wheat

AFLP analysis of near-isogenic lines of the stripe rust resistance gene Yr10 was carried out with 6 PstⅠ- primers and 10 TaqⅠ-primers with the donor parent of Yr10 gene as the check. A total of about 4200 distinguishable bands were amplified, of which 5 were stable. The genetic linkage of the 5 polymorphic DNA fragments with the target gene were tested preliminarily on a segregating F₂ population derived from a cross between the gene donor parent “Moro” and susceptible cultivar “Mingxian 169”. The DNA fragment PT0502 was found closely linked to the Yr10 gene and cloned and sequenced. Based on the sequence specific primers for PCR were designed and synthesized. Genetic linkage analysis with 195 segregating F₂ plants indicated that the genetic distance was 0.5 cM between the main product SC₂₀₀ fragment produced by PCR with the primers and the Yr10 gene. The primers can be used to detect the Yr10 gene quickly, effectively and exactly.     

In vitro assembly of plant tubulin in the absence of microtubule-stabilizing reagents

The assembly of microtubules is essential for physiological functions of microtubules. Addition of microtubule-stabilizing reagents or microtubule “seeds” is usually necessary for plant tubulin assemblyin vitro, which hinders the investigation of plant microtubule dynamics. In the present note, highly purified plant tubulins have been obtained from lily pollen, a non-microtubule-stabilizing reagent or microtubule “seed” system for plant tubulin assembly has been established and the analysis of plant tubulin assembly performed. Experiment results showed that purified tubulin polymerizedin vitro, and a typical microtubule structure was observed with electron microscopy. The kinetics curve of tubulin assembly exhibited typical “parabola”. The presence of taxol significantly altered the character of plant tubulin assembly, including that abnormal microtubules were assembled and the critical concentration for plant tubulin assembly was decreased exceedingly from 3 mg/mL in the absence of taxol to 0.043 mg/mL in ihe presence of taxol.     

Agrobacterium-mediated transformation: state of the art and future prospect

Great progress has been made in recent years in studies on the mechanism ofAgrobacterium-medicated transformation and its application. Many details of the key molecular events within the bacterial cells involved in T-DNA transfer have been elucidated, and it is notable that some plant factors which were elusive before are purified and characterized. Vast kinds of species, which were either recalcitrant to or not included in the host range ofAgrobacterium, can now be transformed by this bacterium, and they include the very important cereal species, gymnosperms, yeast and many filamentous fungi. The simplein vivo transformation of tissue in intact plants and the “agrolistic” methods to transform recalcitrant plants are the two novel technical achievements. Combined with other powerful techniques such as bacterial artificial chromosome, very large DNA fragment can be transformed into the plant genome byAgrobacterium. Further studies will elucidate more plant-encoded factors involved in T-DNA transformation and there is a need to develop more powerfulAgrobacterium-based transformation systems to meet different needs in basic research and crop improvement practice.     

Alp7/TACC is a crucial target in Ran-GTPase-dependent spindle formation in fission yeast

Microtubules are essential intracellular structures involved in several cellular phenomena, including polarity establishment and chromosome segregation. Because the nuclear envelope persists during mitosis (closed mitosis) in fission yeast (Schizosaccharomyces pombe), cytoplasmic microtubules must be reorganized into the spindle in the compartmentalized nucleus on mitotic entry. An ideal mechanism might be to take advantage of an evolutionarily conserved microtubule formation system that uses the Ran-GTPase nuclear transport machinery, but no targets of Ran for spindle formation have been identified in yeast. Here we show that a microtubule-associated protein, Alp7, which forms a complex with Alp14, is a target of Ran in yeast for spindle formation. The Ran-deficient pim1 mutant (pim1-F201S) failed to show mitosis-specific nuclear accumulation of Alp7. Moreover, this mutant exhibited compromised spindle formation and early mitotic delay. Importantly, these defects were suppressed by Alp7 that was artificially targeted to the nucleus by a Ran-independent and importin-alpha-mediated system. Thus, Ran targets Alp7-Alp14 to achieve nuclear spindle formation, and might differentiate its targets depending on whether the organism undergoes closed or open mitosis.     

High precision topographic data on Lop Nor basin＇s Lake ＂Great Ear＂ and the timing of its becoming a dry salt lake

High precision elevation measurements using DGPS were carried out along three representative tran-sects for the ＂Great Ear＂ area, a dry salt lake within the Lop Nor basin. Results indicate that the Lop Nor basin is only 5.2 m deep and its lowest point occurs at the center of the ＂Great Ear＂. In addition, the basin is asymmetric-steeper in the southwest （0.19‰） and gentler in the northeast （0.09‰）. Points along the same ＂Great Ear＂ ring were found to have an identical elevation value, but different when from different ones （lower towards the center）. The spacing of the ＂Great Ear＂ rings was found to be closely related with the surface steepness. The closer the ＂Great Ear＂ rings are spaced, the steeper the ground surface, and vice versa. These findings support the argument that the ＂Great Ear＂ rings are the former shoreline trails left behind by Lop Nor water during the last few episodes of recession towards its total dry up. A comprehensive analysis of the high precision elevation data, historical accounts, aerial and satellite photographs and imagery, and official topographic maps of the study area suggests that the ＂Great Ear＂ area in the Lop Nor basin was incorrectly mapped as being covered by a great body of water on the 1963 topographic maps. A re-interpretation of the 1958 aerial photographs and newer remote sensing imagery indicated that the ＂Great Ear＂ ring structure was already in place in 1958 and it continued to appear on the subsequent remote sensing data without any major changes. It is estimated that lake water in the ＂Great Ear＂ area of the Lop Nor basin disappeared between the late 1930s and early 1940s.     

An only-once-sorting algorithm

This paper provides a new sorting algorithm called “Only-Once-Sorting” algorithm. Using a mathematical formula, this algorithm can put elements in the positions they should be stored only once, then compacts them. The algorithm completes sorting a sequence ofn elements in a calculation time of O(n). Xu Xusong: born in June 1945. Professor. Current research interest is in data structure and algorithm, information systems analysis and design     

Integration of telomere sequences with the draft human genome sequence

Telomeres are the ends of linear eukaryotic chromosomes. To ensure that no large stretches of uncharacterized DNA remain between the ends of the human working draft sequence and the ends of each chromosome, we would need to connect the sequences of the telomeres to the working draft sequence. But telomeres have an unusual DNA sequence composition and organization that makes them particularly difficult to isolate and analyse. Here we use specialized linear yeast artificial chromosome clones, each carrying a large telomere-terminal fragment of human DNA, to integrate most human telomeres with the working draft sequence. Subtelomeric sequence structure appears to vary widely, mainly as a result of large differences in subtelomeric repeat sequence abundance and organization at individual telomeres. Many subtelomeric regions appear to be gene-rich, matching both known and unknown expressed genes. This indicates that human subtelomeric regions are not simply buffers of nonfunctional 'junk DNA' next to the molecular telomere, but are instead functional parts of the expressed genome.     

Study on heterosis of intersubspecies between indica and japonica rice （Oryza sativa L.） using chromosome segment substitution lines

Great heterosis exists in the inter-subspecific crossesbetween indica and japonica rice cultivars[1,2] and the ex-ploitation of this heterosis has long been considered as apromising method to further increase rice yield potential[3].Previous studies indicated that there were large differencesbetween the two subspecies with respect to morphology,isozyme, polymorphism of molecular markers and DNAstructure due to long time genetic differentiation[4,5]. Thosedifferences were identified to be clos…     

Break-induced replication and telomerase-independent telomere maintenance require Pol32

Break-induced replication (BIR) is an efficient homologous recombination process to initiate DNA replication when only one end of a chromosome double-strand break shares homology with a template. BIR is thought to re-establish replication at stalled and broken replication forks and to act at eroding telomeres in cells that lack telomerase in pathways known as 'alternative lengthening of telomeres' (reviewed in refs 2, 6). Here we show that, in haploid budding yeast, Rad51-dependent BIR induced by HO endonuclease requires the lagging strand DNA Polalpha-primase complex as well as Poldelta to initiate new DNA synthesis. Polepsilon is not required for the initial primer extension step of BIR but is required to complete 30 kb of new DNA synthesis. Initiation of BIR also requires the nonessential DNA Poldelta subunit Pol32 primarily through its interaction with another Poldelta subunit, Pol31. HO-induced gene conversion, in which both ends of a double-strand break engage in homologous recombination, does not require Pol32. Pol32 is also required for the recovery of both Rad51-dependent and Rad51-independent survivors in yeast strains lacking telomerase. These results strongly suggest that both types of telomere maintenance pathways occur by recombination-dependent DNA replication. Thus Pol32, dispensable for replication and for gene conversion, is uniquely required for BIR; this finding provides an opening into understanding how DNA replication re-start mechanisms operate in eukaryotes. We also note that Pol32 homologues have been identified both in fission yeast and in metazoans where telomerase-independent survivors with alternative telomere maintenance have also been identified.     

Is there left-handed DNA at the ends of yeast chromosomes?

Tracts of the alternating copolymer poly(dGdT . dCdA) have been observed in a variety of eukaryotes. Such tracts are of particular interest since homopolymers of this sequence can exist in vitro as left-handed Z form DNA. We have found that the yeast Saccharomyces cerevisiae contains at least 30 poly(GT) tracts at dispersed genomic locations. We show here that one subset of these tracts is located at the ends (telomeres) of the yeast chromosome. In addition, we show that poly(dGdT . dCdA) tracts are added to the ends of the extrachromosomal ribosomal DNA molecules of Tetrahymena when cloned in yeast. These data represent the first reported association between a homopolymeric sequence and a chromosome structure.     

Hydrothermal growth of Ni-based coatings on copper substrates

Ni-based coatings were deposited on copper substrates by a hydrothermal approach. The results showed that a Ni-based cellular microstructure was bridged by "fiber-like" products. A high microhardness of Hv 856 was achieved after 400℃ heat treatment, which is nine times that of copper substrates (Hv 95). Nucleation, growth, and fusion of Ni atoms along the linear direction, induced by a linear-type citrate-metal structural "molecule template", led to in-situ growth of Ni-based fibers between cellular microspheres. After 400℃ heat treatment, the precipitation of NiP and Ni₃P hard phases contributed to the high microhardness of Ni-based coatings.     

Optical mapping of a rice BAC clone using restriction endonuclease and imaging with fluorescent microscopy at single molecule level

A method of constructing restriction map by optical mapping and single molecule fluorescent microscopy is described. DNA molecules were aligned and adsorbed on a glass coverslip surface by a modified “molecular combing” technique, and then the surface-immobilized DNAs were cleaved in situ with a restriction endonuclease. Individual DNA molecules digested by the endonuclease EcoRⅠ were observable with fluorescent microscopy. Using optical mapping, a physical map of a rice bacterial artificial chromosome clone was constructed. This method will facilitate genomic mapping and tracing the dynamic process in real time at a single molecule level with fluorescence microscopy.