The Oxytricha trifallax macronuclear genome: a complex eukaryotic genome with 16,000 tiny chromosomes.

The macronuclear genome of the ciliate Oxytricha trifallax displays an extreme and unique eukaryotic genome architecture with extensive genomic variation. During sexual genome development, the expressed, somatic macronuclear genome is whittled down to the genic portion of a small fraction (∼5%) of its precursor "silent" germline micronuclear genome by a process of "unscrambling" and fragmentation. The tiny macronuclear "nanochromosomes" typically encode single, protein-coding genes (a small portion, 10%, encode 2-8 genes), have minimal noncoding regions, and are differentially amplified to an average of ∼2,000 copies. We report the high-quality genome assembly of ∼16,000 complete nanochromosomes (∼50 Mb haploid genome size) that vary from 469 bp to 66 kb long (mean ∼3.2 kb) and encode ∼18,500 genes. Alternative DNA fragmentation processes ∼10% of the nanochromosomes into multiple isoforms that usually encode complete genes. Nucleotide diversity in the macronucleus is very high (SNP heterozygosity is ∼4.0%), suggesting that Oxytricha trifallax may have one of the largest known effective population sizes of eukaryotes. Comparison to other ciliates with nonscrambled genomes and long macronuclear chromosomes (on the order of 100 kb) suggests several candidate proteins that could be involved in genome rearrangement, including domesticated MULE and IS1595-like DDE transposases. The assembly of the highly fragmented Oxytricha macronuclear genome is the first completed genome with such an unusual architecture. This genome sequence provides tantalizing glimpses into novel molecular biology and evolution. For example, Oxytricha maintains tens of millions of telomeres per cell and has also evolved an intriguing expansion of telomere end-binding proteins. In conjunction with the micronuclear genome in progress, the O. trifallax macronuclear genome will provide an invaluable resource for investigating programmed genome rearrangements, complementing studies of rearrangements arising during evolution and disease.

Defining the regions of homology between human chromosome 16p12--13 and mouse chromosomes

In this study, the evolutionary relationship between human chromosome 16p12-p13 and mouse chromosomes was investigated by determining the order of marker loci in the region and then identifying the chromosomal locations of the homologous loci in mice. Eighteen genes from human 16 were mapped to fifteen subchromosomal regions by a variety of mapping approaches.^ Thirteen of the genes were mapped in the mouse. Linkage analysis with backcross mice and segregation analysis in a mouse - Chinese Hamster Ovary (CHO) somatic cell hybrid panel informative for different regions of mouse genome were used. The results assigned the thirteen genes to three different mouse chromosomes.^ A group of six genes on mouse 16 was found to be closely linked to Scid. The order of Myh11 and Mrp remains ambiguous since no recombination was detected in backcross analysis. Their relative position in human is also uncertain since they were shown to be very close to each other. For the other mouse loci, an unambiguous gene order could be determined and was found to be identical to that in human. Therefore, they comprise a new conserved linkage group between the two species. The orientation of the group was inverted relative to the centromeres, i.e. the proximal loci in one species become distal in another. The size of the group was estimated to be from 4.4 to 8 Mb and 10 to 32 cM in human. In mouse, it was about 21 cM in the backcross analysis. The two boundaries of the conserved linkage were defined within a 1 Mb range. It is now possible to predict the locations of mouse homologs for some human disease genes based on their locations on human 16p.^ The six human 16p genes that map to MMU7 showed a different gene order in mouse than in human. No recombination was found between Crym and Umod while Crym was distal to D16S79A and proximal to D16S92. The location of Stp and Cdr2 with respect to the above four loci was not determined since they were not mapped in the same set of backcross mice. These genes greatly expanded an existing conserved synteny group between the human 16p12-p13 region and the MMU7. It now consists of eleven loci that span a region of probably more than 10 Mb in human. The gene order derived from this study provided further evidence for chromosomal rearrangements within the conserved synteny. (Abstract shortened by UMI.) ^

PREMATURE CONDENSATION OF SPERMATOGENIC CELL CHROMOSOMES

The technique of premature chromosome condensation (PCC) has been used primarily to study interphase chromosomes of somatic cells. In this study, mitotic cells were fused to cells from the mouse testes to examine the chromosomes of germ cells. The testes contain various types of cells, both germinal and nongerminal. In these initial studies, four types of PCC morphologies were observed. Chromosome morphology of the PCC and labeling experiments demonstrated the mouse cell origin of various PCC. Attempts were next made to determine the cell types producing the PCC. Spermatogonia, diplotene spermatocytes, secondary spermatocytes and round spermatids are proposed to be the origin of the PCC morphologies. Some PCC could be banded by G and C banding techniques and the mouse chromosomes identified.^ Studies were subsequently undertaken to evaluate this technique as a method of evaluating damage to germ cells. Testicular cells from irradiated mice were fused to mitotic cells and the PCC examined. Both round spermatids and secondary spermatocytes exhibited chromosome damage in the form of chromatid breaks. A linear correlation was found between the dose of irradiation and the number of breaks per cell. This technique may develop into a useful method for evaluating the clastogenic effect of agents on the germ cells. ^

Morphologic characteristics, location, and associated complications of maxillary and mandibular supernumerary teeth as evaluated using cone beam computed tomography.

OBJECTIVES To evaluate the location and morphologic characteristics of supernumerary teeth and to assess the frequency and extent of root resorption of adjacent teeth using cone beam computed tomography (CBCT). MATERIALS AND METHODS CBCT scans of 82 patients with supernumerary teeth in the maxilla and mandible were evaluated by two orthodontists independently. Data regarding the type, shape, and three-dimensional (3D) location of the supernumeraries including the frequency and extent of root resorption of adjacent teeth were recorded and evaluated for possible associations. RESULTS The study comprised a total of 101 supernumerary teeth. Most of the patients (80.5 per cent) exhibited one single supernumerary tooth, while 15.8 per cent had two and 3.7 per cent had three supernumeraries. Males were affected more than females with a ratio of 1.65:1. Mesiodentes were the most frequently diagnosed type of supernumerary teeth (48.52 per cent), followed by supernumerary premolars (23.76 per cent) and lateral incisors (18.81 per cent). Supernumeraries were most commonly conical in shape (42.6 per cent) with a normal or inclined vertical position (61.4 per cent). Root resorption of adjacent teeth was detected for 22.8 per cent of the supernumerary teeth, most frequently for supernumerary premolars. There was a significant association between root resorption of adjacent teeth and type and shape of tooth. Interrater agreement for the measurements performed showed kappa values ranging from 0.55 to 1 with a kappa value of 1 for type and shape of the supernumerary teeth. CONCLUSIONS CBCT provides 3D information about location and shape of supernumerary teeth as well as prevalence and degree of root resorption of neighbouring teeth with moderate to high interrater correlation.

THE MICROCELL-MEDIATED TRANSFER OF SINGLE HUMAN CHROMOSOMES INTO RECIPIENT MOUSE CELLS

Microcell-mediated chromosome transfer is a method of gene transfer which allows for the introduction of single or small groups of intact chromosomes into recipient host cells. Microcell transfer was first performed by Fournier and Ruddle using rodent microcells and various recipient cells. Expansion of this technology to include the transfer of normal human genetic material has been hindered because large micronucleate populations from diploid human cells have been unobtainable. This dissertation research describes, however, the methods for production of micronuclei in 40-60% of normal human fibroblasts. Once micronucleate cells were obtained, they were enucleated by centrifugation in the presence of Cytochalasin B; the microcells were then purified and fused to recipient mouse (LMTK('-)) cells using a new fusion protocol employing polyethylene glycol containing phytohemagglutinin. Microcell clones were isolated from the HAT selection system. Alkaline Giemsa staining performed on these hybrids indicated the presence of a single human chromosome in each of seven microcell clones from three separate experiments. That chromosome was further identified by G banding analysis to be human chromosome #17, which codes for thymidine kinase. The time course for production of these hybrids from fusion to karyotypic analysis was 6 weeks. The viability of the transferred human genetic material was assessed by electrophoretic isozyme analysis.^ Subsequent experiments were performed in an attempt to optimize the transfer frequency for the thymidine kinase gene using this system. Results indicated that the frequency could be increased from < 1 x 10('-6) in initial experiments to 2 x 10('-5) in the latest experiment. Analyses were also conducted to determine the number of chromosomes per isolated microcell as well as to investigate the stability of the transferred human chromosome in the mouse genome. ^

CYTOLOGICAL, ULTRASTRUCTURAL, AND BIOCHEMICAL STUDIES OF THE STRUCTURE OF PREMATURELY CONDENSED CHROMOSOMES

The phenomenon of premature chromosome condensation, resulting from fusion between mitotic and interphase cells, includes dissolution of the interphase nuclear framework, thus allowing a direct visualization of interphase chromosomes. Light microscope morphology of prematurely condensed chromosomes (PCC) from synchronized HeLa cells supports the model of an interphase "chromosome condensation cycle". PCC are increasingly attenuated as cells progress through G(,1). A maximum degree of decondensation is observed at active sites of DNA replication during S phase, and a condensed morphology is rapidly resumed following completion of replication of a chromosome segment.^ To permit ultrastructural and biochemical studies of PCC, a procedure was developed to induce premature chromosome condensation at high frequency. This was achieved by polyethylene glycol (PEG)-mediated fusion of a dense monolayer of mitotic and interphase cells induced by centrifugation onto lectin-coated culture dishes. Using this method, PCC induction frequencies of 60-90% are routinely obtained.^ Scanning electron microscope analysis of PCC spreads revealed that the extension of PCC during progression through G(,1) is accompanied by a transition of the basic 30 nm chromatin fiber from tightly packed looping fibers to extended longitudinal fibers. Sites of active DNA replication is S-PCC were indicated to be organized a single longitudinal fibers. Following replication of a chromosome segment, a rapid reorganization from the extended longitudinal fiber to packed looping fibers occurs. The postreplication maturation process appears to include the assembly of a chromosome core consisting of multiple longitudinal fibers.^ The role of histone H1 phosphorylation in PCC formation was investigated by acidurea polyacrylamide gel electrophoresis of total histone extracted from metaphase chromosomes and PCC following high frequency fusion. This investigation failed to demonstrate an extensive phosphorylation of H1 associated with PCC formation. However, significant dephosphorylation of superphosphorylated metaphase chromosome H1 was observed, indicating that interphase H1-phosphatase activity is dominant over metaphase H1 kinase activity. These observations provide evidence against models suggesting a role for H1 superphosphorylation in triggering mitotic condensation of chromosomes. ^

IDENTIFICATION AND ANALYSIS OF A NOVEL ROLE FOR THE TOUSLED-LIKE KINASE IN REGULATING MITOTIC SPINDLE DYNAMICS

Deregulation of kinase activity is one example of how cells become cancerous by evading evolutionary constraints. The Tousled kinase (Tsl) was initially identified in Arabidopsis thaliana as a developmentally important kinase. There are two mammalian orthologues of Tsl and one orthologue in C. elegans, TLK-1, which is essential for embryonic viability and germ cell development. Depletion of TLK-1 leads to embryonic arrest large, distended nuclei, and ultimately embryonic lethality. Prior to terminal arrest, TLK-1-depleted embryos undergo aberrant mitoses characterized by poor metaphase chromosome alignment, delayed mitotic progression, lagging chromosomes, and supernumerary centrosomes. I discovered an unanticipated requirement for TLK-1 in mitotic spindle assembly and positioning. Normally, in the newly-fertilized zygote (P0) the maternal pronucleus migrates toward the paternal pronucleus at the posterior end of the embryo. After pronuclear meeting, the pronuclear-centrosome complex rotates 90° during centration to align on the anteroposterior axis followed by nuclear envelope breakdown (NEBD). However, in TLK-1-depleted P0 embryos, the centrosome-pronuclear complex rotation is significantly delayed with respect to NEBD and chromosome congression, Additionally, centrosome positions over time in tlk-1(RNAi) early embryos revealed a defect in posterior centrosome positioning during spindle-pronuclear centration, and 4D analysis of centrosome positions and movement in newly fertilized embryos showed aberrant centrosome dynamics in TLK-1-depleted embryos. Several mechanisms contribute to spindle rotation, one of which is the anchoring of astral microtubules to the cell cortex. Attachment of these microtubules to the cortices is thought to confer the necessary stability and forces in order to rotate the centrosome-pronuclear complex in a timely fashion. Analysis of a microtubule end-binding protein revealed that TLK-1-depleted embryos exhibit a more stochastic distribution of microtubule growth toward the cell cortices, and the types of microtubule attachments appear to differ from wild-type embryos. Additionally, fewer astral microtubules are in the vicinity of the cell cortex, thus suggesting that the delayed spindle rotation could be in part due to a lack of appropriate microtubule attachments to the cell cortex. Together with recently published biochemical data revealing the Tousled-like kinases associate with components of the dynein microtubule motor complex in humans, these data suggest that Tousled-like kinases play an important role in mitotic spindle assembly and positioning.

Detection and pattern recognition applied to leaves and chromosomes

The Project you are about to see it is based on the technologies used on object detection and recognition, especially on leaves and chromosomes. To do so, this document contains the typical parts of a scientific paper, as it is what it is. It is composed by an Abstract, an Introduction, points that have to do with the investigation area, future work, conclusions and references used for the elaboration of the document. The Abstract talks about what are we going to find in this paper, which is technologies employed on pattern detection and recognition for leaves and chromosomes and the jobs that are already made for cataloguing these objects. In the introduction detection and recognition meanings are explained. This is necessary as many papers get confused with these terms, specially the ones talking about chromosomes. Detecting an object is gathering the parts of the image that are useful and eliminating the useless parts. Summarizing, detection would be recognizing the objects borders. When talking about recognition, we are talking about the computers or the machines process, which says what kind of object we are handling. Afterwards we face a compilation of the most used technologies in object detection in general. There are two main groups on this category: Based on derivatives of images and based on ASIFT points. The ones that are based on derivatives of images have in common that convolving them with a previously created matrix does the treatment of them. This is done for detecting borders on the images, which are changes on the intensity of the pixels. Within these technologies we face two groups: Gradian based, which search for maximums and minimums on the pixels intensity as they only use the first derivative. The Laplacian based methods search for zeros on the pixels intensity as they use the second derivative. Depending on the level of details that we want to use on the final result, we will choose one option or the other, because, as its logic, if we used Gradian based methods, the computer will consume less resources and less time as there are less operations, but the quality will be worse. On the other hand, if we use the Laplacian based methods we will need more time and resources as they require more operations, but we will have a much better quality result. After explaining all the derivative based methods, we take a look on the different algorithms that are available for both groups. The other big group of technologies for object recognition is the one based on ASIFT points, which are based on 6 image parameters and compare them with another image taking under consideration these parameters. These methods disadvantage, for our future purposes, is that it is only valid for one single object. So if we are going to recognize two different leaves, even though if they refer to the same specie, we are not going to be able to recognize them with this method. It is important to mention these types of technologies as we are talking about recognition methods in general. At the end of the chapter we can see a comparison with pros and cons of all technologies that are employed. Firstly comparing them separately and then comparing them all together, based on our purposes. Recognition techniques, which are the next chapter, are not really vast as, even though there are general steps for doing object recognition, every single object that has to be recognized has its own method as the are different. This is why there is not a general method that we can specify on this chapter. We now move on into leaf detection techniques on computers. Now we will use the technique explained above based on the image derivatives. Next step will be to turn the leaf into several parameters. Depending on the document that you are referring to, there will be more or less parameters. Some papers recommend to divide the leaf into 3 main features (shape, dent and vein] and doing mathematical operations with them we can get up to 16 secondary features. Next proposition is dividing the leaf into 5 main features (Diameter, physiological length, physiological width, area and perimeter] and from those, extract 12 secondary features. This second alternative is the most used so it is the one that is going to be the reference. Following in to leaf recognition, we are based on a paper that provides a source code that, clicking on both leaf ends, it automatically tells to which specie belongs the leaf that we are trying to recognize. To do so, it only requires having a database. On the tests that have been made by the document, they assure us a 90.312% of accuracy over 320 total tests (32 plants on the database and 10 tests per specie]. Next chapter talks about chromosome detection, where we shall pass the metaphasis plate, where the chromosomes are disorganized, into the karyotype plate, which is the usual view of the 23 chromosomes ordered by number. There are two types of techniques to do this step: the skeletonization process and swiping angles. Skeletonization progress consists on suppressing the inside pixels of the chromosome to just stay with the silhouette. This method is really similar to the ones based on the derivatives of the image but the difference is that it doesnt detect the borders but the interior of the chromosome. Second technique consists of swiping angles from the beginning of the chromosome and, taking under consideration, that on a single chromosome we cannot have more than an X angle, it detects the various regions of the chromosomes. Once the karyotype plate is defined, we continue with chromosome recognition. To do so, there is a technique based on the banding that chromosomes have (grey scale bands] that make them unique. The program then detects the longitudinal axis of the chromosome and reconstructs the band profiles. Then the computer is able to recognize this chromosome. Concerning the future work, we generally have to independent techniques that dont reunite detection and recognition, so our main focus would be to prepare a program that gathers both techniques. On the leaf matter we have seen that, detection and recognition, have a link as both share the option of dividing the leaf into 5 main features. The work that would have to be done is to create an algorithm that linked both methods, as in the program, which recognizes leaves, it has to be clicked both leaf ends so it is not an automatic algorithm. On the chromosome side, we should create an algorithm that searches for the beginning of the chromosome and then start to swipe angles, to later give the parameters to the program that searches for the band profiles. Finally, on the summary, we explain why this type of investigation is needed, and that is because with global warming, lots of species (animals and plants] are beginning to extinguish. That is the reason why a big database, which gathers all the possible species, is needed. For recognizing animal species, we just only have to have the 23 chromosomes. While recognizing a plant, there are several ways of doing it, but the easiest way to input a computer is to scan the leaf of the plant. RESUMEN. El proyecto que se puede ver a continuación trata sobre las tecnologías empleadas en la detección y reconocimiento de objetos, especialmente de hojas y cromosomas. Para ello, este documento contiene las partes típicas de un paper de investigación, puesto que es de lo que se trata. Así, estará compuesto de Abstract, Introducción, diversos puntos que tengan que ver con el área a investigar, trabajo futuro, conclusiones y biografía utilizada para la realización del documento. Así, el Abstract nos cuenta qué vamos a poder encontrar en este paper, que no es ni más ni menos que las tecnologías empleadas en el reconocimiento y detección de patrones en hojas y cromosomas y qué trabajos hay existentes para catalogar a estos objetos. En la introducción se explican los conceptos de qué es la detección y qué es el reconocimiento. Esto es necesario ya que muchos papers científicos, especialmente los que hablan de cromosomas, confunden estos dos términos que no podían ser más sencillos. Por un lado tendríamos la detección del objeto, que sería simplemente coger las partes que nos interesasen de la imagen y eliminar aquellas partes que no nos fueran útiles para un futuro. Resumiendo, sería reconocer los bordes del objeto de estudio. Cuando hablamos de reconocimiento, estamos refiriéndonos al proceso que tiene el ordenador, o la máquina, para decir qué clase de objeto estamos tratando. Seguidamente nos encontramos con un recopilatorio de las tecnologías más utilizadas para la detección de objetos, en general. Aquí nos encontraríamos con dos grandes grupos de tecnologías: Las basadas en las derivadas de imágenes y las basadas en los puntos ASIFT. El grupo de tecnologías basadas en derivadas de imágenes tienen en común que hay que tratar a las imágenes mediante una convolución con una matriz creada previamente. Esto se hace para detectar bordes en las imágenes que son básicamente cambios en la intensidad de los píxeles. Dentro de estas tecnologías nos encontramos con dos grupos: Los basados en gradientes, los cuales buscan máximos y mínimos de intensidad en la imagen puesto que sólo utilizan la primera derivada; y los Laplacianos, los cuales buscan ceros en la intensidad de los píxeles puesto que estos utilizan la segunda derivada de la imagen. Dependiendo del nivel de detalles que queramos utilizar en el resultado final nos decantaremos por un método u otro puesto que, como es lógico, si utilizamos los basados en el gradiente habrá menos operaciones por lo que consumirá más tiempo y recursos pero por la contra tendremos menos calidad de imagen. Y al revés pasa con los Laplacianos, puesto que necesitan más operaciones y recursos pero tendrán un resultado final con mejor calidad. Después de explicar los tipos de operadores que hay, se hace un recorrido explicando los distintos tipos de algoritmos que hay en cada uno de los grupos. El otro gran grupo de tecnologías para el reconocimiento de objetos son los basados en puntos ASIFT, los cuales se basan en 6 parámetros de la imagen y la comparan con otra imagen teniendo en cuenta dichos parámetros. La desventaja de este método, para nuestros propósitos futuros, es que sólo es valido para un objeto en concreto. Por lo que si vamos a reconocer dos hojas diferentes, aunque sean de la misma especie, no vamos a poder reconocerlas mediante este método. Aún así es importante explicar este tipo de tecnologías puesto que estamos hablando de técnicas de reconocimiento en general. Al final del capítulo podremos ver una comparación con los pros y las contras de todas las tecnologías empleadas. Primeramente comparándolas de forma separada y, finalmente, compararemos todos los métodos existentes en base a nuestros propósitos. Las técnicas de reconocimiento, el siguiente apartado, no es muy extenso puesto que, aunque haya pasos generales para el reconocimiento de objetos, cada objeto a reconocer es distinto por lo que no hay un método específico que se pueda generalizar. Pasamos ahora a las técnicas de detección de hojas mediante ordenador. Aquí usaremos la técnica explicada previamente explicada basada en las derivadas de las imágenes. La continuación de este paso sería diseccionar la hoja en diversos parámetros. Dependiendo de la fuente a la que se consulte pueden haber más o menos parámetros. Unos documentos aconsejan dividir la morfología de la hoja en 3 parámetros principales (Forma, Dentina y ramificación] y derivando de dichos parámetros convertirlos a 16 parámetros secundarios. La otra propuesta es dividir la morfología de la hoja en 5 parámetros principales (Diámetro, longitud fisiológica, anchura fisiológica, área y perímetro] y de ahí extraer 12 parámetros secundarios. Esta segunda propuesta es la más utilizada de todas por lo que es la que se utilizará. Pasamos al reconocimiento de hojas, en la cual nos hemos basado en un documento que provee un código fuente que cucando en los dos extremos de la hoja automáticamente nos dice a qué especie pertenece la hoja que estamos intentando reconocer. Para ello sólo hay que formar una base de datos. En los test realizados por el citado documento, nos aseguran que tiene un índice de acierto del 90.312% en 320 test en total (32 plantas insertadas en la base de datos por 10 test que se han realizado por cada una de las especies]. El siguiente apartado trata de la detección de cromosomas, en el cual se debe de pasar de la célula metafásica, donde los cromosomas están desorganizados, al cariotipo, que es como solemos ver los 23 cromosomas de forma ordenada. Hay dos tipos de técnicas para realizar este paso: Por el proceso de esquelotonización y barriendo ángulos. El proceso de esqueletonización consiste en eliminar los píxeles del interior del cromosoma para quedarse con su silueta; Este proceso es similar a los métodos de derivación de los píxeles pero se diferencia en que no detecta bordes si no que detecta el interior de los cromosomas. La segunda técnica consiste en ir barriendo ángulos desde el principio del cromosoma y teniendo en cuenta que un cromosoma no puede doblarse más de X grados detecta las diversas regiones de los cromosomas. Una vez tengamos el cariotipo, se continua con el reconocimiento de cromosomas. Para ello existe una técnica basada en las bandas de blancos y negros que tienen los cromosomas y que son las que los hacen únicos. Para ello el programa detecta los ejes longitudinales del cromosoma y reconstruye los perfiles de las bandas que posee el cromosoma y que lo identifican como único. En cuanto al trabajo que se podría desempeñar en el futuro, tenemos por lo general dos técnicas independientes que no unen la detección con el reconocimiento por lo que se habría de preparar un programa que uniese estas dos técnicas. Respecto a las hojas hemos visto que ambos métodos, detección y reconocimiento, están vinculados debido a que ambos comparten la opinión de dividir las hojas en 5 parámetros principales. El trabajo que habría que realizar sería el de crear un algoritmo que conectase a ambos ya que en el programa de reconocimiento se debe clicar a los dos extremos de la hoja por lo que no es una tarea automática. En cuanto a los cromosomas, se debería de crear un algoritmo que busque el inicio del cromosoma y entonces empiece a barrer ángulos para después poder dárselo al programa que busca los perfiles de bandas de los cromosomas. Finalmente, en el resumen se explica el por qué hace falta este tipo de investigación, esto es que con el calentamiento global, muchas de las especies (tanto animales como plantas] se están empezando a extinguir. Es por ello que se necesitará una base de datos que contemple todas las posibles especies tanto del reino animal como del reino vegetal. Para reconocer a una especie animal, simplemente bastará con tener sus 23 cromosomas; mientras que para reconocer a una especie vegetal, existen diversas formas. Aunque la más sencilla de todas es contar con la hoja de la especie puesto que es el elemento más fácil de escanear e introducir en el ordenador.

Segregation of viral plasmids depends on tethering to chromosomes and is regulated by phosphorylation

Eukaryotic viruses can maintain latency in dividing cells as extrachromosomal nuclear plasmids. Segregation and nuclear retention of DNA is, therefore, a key issue in retaining copy number. The E2 enhancer protein of the papillomaviruses is required for viral DNA replication and transcription. Viral mutants that prevent phosphorylation of the bovine papillomavirus type 1 (BPV) E2 protein are transformation-defective, despite normal viral gene expression and replication function. Cell colonies harboring such mutants show sectoring of viral DNA and are unable to maintain the episome. We find that transforming viral DNA attaches to mitotic chromosomes, in contrast to the mutant genome encoding the E2 phosphorylation mutant. Second-site suppressor mutations were uncovered in both E1 and E2 genes that allow for transformation, maintenance, and chromosomal attachment. E2 protein was also found to colocalize to mitotic chromosomes, whereas the mutant did not, suggesting a direct role for E2 in viral attachment to chromosomes. Such viral hitch-hiking onto cellular chromosomes is likely to provide a general mechanism for maintaining nuclear plasmids.