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2.
Yasutaka Kakui Tomonari Sunaga Kunio Arai James Dodgson Liang Ji Attila Csikász-Nagy Rafael Carazo-Salas Masamitsu Sato 《Open biology》2015,5(6)
Integration of an external gene into a fission yeast chromosome is useful to investigate the effect of the gene product. An easy way to knock-in a gene construct is use of an integration plasmid, which can be targeted and inserted to a chromosome through homologous recombination. Despite the advantage of integration, construction of integration plasmids is energy- and time-consuming, because there is no systematic library of integration plasmids with various promoters, fluorescent protein tags, terminators and selection markers; therefore, researchers are often forced to make appropriate ones through multiple rounds of cloning procedures. Here, we establish materials and methods to easily construct integration plasmids. We introduce a convenient cloning system based on Golden Gate DNA shuffling, which enables the connection of multiple DNA fragments at once: any kind of promoters and terminators, the gene of interest, in combination with any fluorescent protein tag genes and any selection markers. Each of those DNA fragments, called a ‘module’, can be tandemly ligated in the order we desire in a single reaction, which yields a circular plasmid in a one-step manner. The resulting plasmids can be integrated through standard methods for transformation. Thus, these materials and methods help easy construction of knock-in strains, and this will further increase the value of fission yeast as a model organism. 相似文献
3.
F-actin organization in the tip-growing cells of fern protonematawas investigated by rhodamine-phalloidin staining in two species:Adiantum capillus-veneris and Pteris vittata. Circular arrangementof cortical F-actin was found around the subapical region ofprotonemal cells in both species. In rhizoids, such structureswere absent and the axial filaments appeared to fan out fromthe tip. (Received May 22, 1989; Accepted September 6, 1989) 相似文献
4.
Masamitsu Wada 《Journal of plant research》1988,101(4):519-528
Chloroplast proliferation was investigated inAdiantum protonemata growing under continuous red light. Cell division is absent when cells are grown under red light. The chloroplast
number increases as the cell length increases, therefore the chloroplasts divide in the absence of cell division. Chloroplasts
in the basal part of the filamentous protonemal cell migrate gradually toward the cell apex, but there is no large net migration
from the tip to the base or vice versa, indicating that chloroplast division takes place in the apical part of the protonemata.
Chloroplast number in the apical 100 μm was maintained at about 200 during cell growth at least over eight days. The chloroplasts
were either dumbbell- or ellipsoid-shaped. Dumbbell-shaped chloroplasts are abundant everywhere in a protonema, ranging from
30 to 50% of the total chloroplasts. The dumbbell-shaped chloroplasts attached to or very close to the plasma membrane seem
to be the ones that are dividing but the dumbbell-shaped ones in the other regions do not divide. These data support the hypothesis
that a signal from the plasma membrane induces the dumbbell-shaped chloroplasts to divide. 相似文献
5.
Mitsuaki Kameko Miho Ichikawa Tsutomu Katsuyama Masamitsu Kanai Michimasa Kato Taiji Akamatsu 《The Histochemical journal》1986,18(4):164-168
Summary This study was undertaken, employing the immunoenzyme method, to confirm the presence of retinol-binding protein in human pancreatic islets, and to compare its distribution with that of prealbumin, insulin, glucagon, somatostatin and pancreatic polypeptide. It was found that most islet cells contained retinol-binding protein, although centrally located cells showed stronger reactivity than those in the peripheral region. The distribution of each of the five polypeptides differed from that of retinolbinding protein, indicating that these peptides did not cross-react with anti-retinol-binding protein antibody. Islet cells which contained prealbumin, on the other hand, were mostly classified as A cells. Further studies are necessary to confirm whether the islet cells produce retinol-binding protein or only store it. 相似文献
6.
Abstract The adherence of Fusobacterium necrophorum to the surface of bovine hepatic cells was investigated. Hemagglutinating strain VPI2891 adhered well to the cell membranes, whereas non-hemagglutinating strain S-45 adhered less well. The adherence of the former strain was strongly inhibited by the homologous anti-hemagglutinin serum. Immunofluorescence study revealed that the purified hemagglutinin bound to the membranes of the hepatic cells. These findings suggest that the adherence of F. necrophorum to the bovine hepatic cells is mediated by the bacterial hemagglutinin and has pathogenic importance in bovine necrobacillosis. 相似文献
7.
Measuring the ratio of the number of photooriented chloroplaststo the total number of chloroplasts, we found that photoorientationof chloroplasts in protonemata of the fern Adiantum capillus-veneriscould be induced by brief irradiation with polarized red light.After irradiation with red light (R) of 3 or 10 min, orientationalmovement was detected as early as 10 min after the irradiation;it continued during the subsequent dark period for 3060min, after which chloroplasts gradually dispersed again. WhenR-treated protonemata were irradiated briefly with a second10-min pulse of R, 60 min after the onset of the first irradiation,the orientational response of chloroplasts was again observed.Typical red/far-red photoreversibility was apparent in the response,indicating the involvement of phytochrome. By contrast, irradiationwith polarized blue light for 10 min was ineffective, whileirradiation with blue light (B) at the same fluence for a longerperiod of time clearly induced the photoorientation of chloroplasts.It is likely that longterm irradiation is necessary for theresponse mediated by a blue-light receptor. When protonemata were irradiated with far-red light (FR) immediatelyafter R or after a subsequent dark period of 10 min, the magnitudeof the orientational response was smaller and chloroplasts dispersedmore quickly than those exposed to R alone. When FR was appliedat 50 min, when the response to R had reached the maximum level,chloroplasts again dispersed rapidly to their dark positions.These results indicate that PFR not only induces the photoorientationmovement of chloroplasts but also fixes the chloroplasts atthe sites to which they have moved as a result of photoorientation. (Received June 2, 1993; Accepted January 11, 1994) 相似文献
8.
Organization of microtubules (MTs) in relation to the behavior of nuclei was examined in dividing binucleate cells ofAdiantum capillus-veneris L. To induce binucleate cells, caffeine, an inhibitor of formation of the cell plate, was applied at 4 mM to synchronously
dividing protonemal cells during cytokinesis (Murata and Wada 1993). Formation of the preprophase band (PPB) during the next
cell cycle was examined in non-centrifuged and centrifuged cells. The two nuclei were separated or associated with one another
in both non-centrifuged and centrifuged cells, although the location of the nuclei in the cylindrical protonemal cells was
different (Murata and Wada 1993). Irrespective of centrifugation, a single PPB was formed around the nuclei in cells with
associated nuclei. Two PPBs were formed in cells with separated nuclei in centrifuged cells. Patterns of mitosis and cytokinesis
varied, depending on the location of the PPB and the distribution of the nuclei. The role of the nucleus in formation of the
PPB is discussed. 相似文献
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