Abstract:
:To construct recombinant retroviruses with only a single active promoter, we introduced point mutations into the TATA box region of the 3'-LTR, and successfully obtained high-titer virus with sufficient self-inactivating activity. However, the viral titer could not be determined by the number of G418 resistant colonies since the neomycin resistance gene was under 5'-LTR control, because of inactivation of the selection marker in target glioma cells. To overcome this problem, we constructed PCR primers with homology to a gene under the control of the internal promoter of recombinant retrovirus, and to retrovirus-specific sequences. There was good correlation between the amount of PCR-amplified product and the number of colony forming units when glioma cells were transduced with the retroviruses containing both the neomycin resistance gene and the HTK gene. Amplified PCR products quantitated by densitometry after glioma cells were transduced with SIV retrovirus vectors, and there was good correlation between density and sensitivity to GCV following transduction. Therefore, detection of HTK PCR products from glioma cells transduced with HTK-bearing retroviruses is useful for determining the appropriate packaging cell for efficient production of viral particles. This detection system is especially useful for isolating high titer clones producing SIV-type retroviruses.
journal_name
Cell Struct Functjournal_title
Cell structure and functionauthors
Miyao Y,Shimizu K,Tamura M,Yamada M,Tamura K,Nakahira K,Kuriyama S,Hayakawa T,Ikenaka Kdoi
10.1247/csf.20.177subject
Has Abstractpub_date
1995-04-01 00:00:00pages
177-83issue
2eissn
0386-7196issn
1347-3700journal_volume
20pub_type
杂志文章abstract::Spindle pole bodies (SPBs) attached to nuclei were isolated from Saccharomyces cerevisiae; they nucleated microtubules to form asters in vitro. This aster-forming activity was stable in 30% dimethylsulfoxide or 10% 2-propanol and could be solubilized with KCl. The KCl extract contained many protein components, which a...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.15.221
更新日期:1990-08-01 00:00:00
abstract::Immunoblots of proteins extracted from the skin of a small viviparous fish (Xiphophorus) showed that a monoclonal antibody against human urokinase recognizes multiple molecular weight species of antigens. The immunoaffinity-purified antigens had serine-protease activity for the hydrolysis of a chromogenic substrate an...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.12.11
更新日期:1987-02-01 00:00:00
abstract::A voltage-dependent and Ca2(+)-activated cation channel recently found in the vacuolar membrane of the yeast Saccharomyces cerevisiae was incorporated into planar lipid bilayers and further characterized in macroscopic and single channel levels. Single channel conductances for various cations were in the order: NH4+ g...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.14.659
更新日期:1989-12-01 00:00:00
abstract::Tobacco BY-2 cells were synchronized by an aphidicolin treatment, and their beta-tubulin isoforms and their mRNA were analyzed by Western, Northern and dot blottings. The relative ratio of the beta-tubulin isoforms changed with the progress of cell cycle stage. By Northern blot hybridization of poly(A)+RNAs with a clo...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.16.489
更新日期:1991-12-01 00:00:00
abstract::It was recently reported that intravenous administration of phorbol 12-myristate 13-acetate (PMA) showed a therapeutic effect in myelocytic leukemia patients. However, we previously observed that, in serum-free conditions, polymorphonuclear leucocytes (PMNs) were killed rapidly by exposure to PMA, suggesting the possi...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.26.235
更新日期:2001-08-01 00:00:00
abstract::Contact reaction is the morphological expression of allogeneic recognition by hemocytes in the ascidian, Halocynthia roretzi; namely they undergo an exocytotic burst upon contact with non-self (allogeneic) hemocytes in vitro. We have found that contact reaction is accompanied by a burst of cyanide-insensitive oxygen c...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.20.81
更新日期:1995-02-01 00:00:00
abstract::Retinoids (retinoic acid and its analogs) are widely involved in the control of cell proliferation, cell differentiation and embryonic development. A series of potent and novel synthetic retinoids named retinobenzoic acids has been developed. Retinobenzoic acids have proven to be useful tools in the investigation of t...
journal_title:Cell structure and function
pub_type: 杂志文章,评审
doi:10.1247/csf.16.113
更新日期:1991-04-01 00:00:00
abstract::The transcription factor ATF6 is constitutively synthesized as a type II transmembrane protein embedded in the endoplasmic reticulum (ER). When unfolded proteins accumulate in the ER, ATF6 senses such ER stress via an as yet undetermined mechanism and relocates to the Golgi apparatus where it is cleaved by sequential ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.10010
更新日期:2011-01-01 00:00:00
abstract::The cell cycle of HeLa S3 cells synchronized by hydroxyurea was investigated by flow cytometry. Metachromatic fluorochrome acridine orange was used to strain the DNA and RNA of the cells differentially. Periodic changes in the cellular DNA and RNA contents were observed through five cell cycles. The G1 and S phases of...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.8.303
更新日期:1983-09-01 00:00:00
abstract::The fine structural localization of albumin in rat liver parenchymal cells was determined by an improved immunocytochemical method and serial sectioning. Albumin in the secretory apparatus of the parenchymal cells was present in segments of the rough endoplasmic reticulum, interrupted with negative segments, in transp...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.12.251
更新日期:1987-06-01 00:00:00
abstract::Mouse FM3A cells propagated well in serum-free medium containing 0.5% serum albumin and 1 microgram of insulin/ml. The vitamin B12 (B12) requirement of the cells depended on the population density. This requirement disappeared when a sufficiently large cell population was present. A combination of 1-100 ng of B12/ml a...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.11.9
更新日期:1986-03-01 00:00:00
abstract::alpha-1,3-Fucosyltransferase (Fuc T IV) cDNA was placed under the control of beta-actin, cytomegalovirus enhancer/promoter and transfected into embryonic stem cells. The transfected cell clones with integrated cDNA (positive clones) differentiated more efficiently into myocardial cell clones without integrated cDNA (n...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.22.247
更新日期:1997-04-01 00:00:00
abstract::The Golgi stress response is a homeostatic mechanism that controls the capacity of the Golgi apparatus in accordance with cellular demands. When the capacity of the Golgi apparatus becomes insufficient (Golgi stress), transcription levels of Golgi-related genes encoding glycosylation enzymes, a Golgi structural protei...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.16005
更新日期:2016-07-30 00:00:00
abstract::Murine tracheal epithelial cell lines, TM01 and TM02-3, were established from a primary culture of tracheal cells of adult transgenic mice bearing a temperature-sensitive simian virus (SV40) large T-antigen gene. Both TM01 and TM02-3 cells, which grew until confluent monolayers were formed, maintained tight contact wi...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.23.119
更新日期:1998-06-01 00:00:00
abstract::The localization of mRNA in neuronal dendrites plays a role in both locally and temporally regulated protein synthesis, which is required for certain forms of synaptic plasticity. RNA granules constitute a dendritic mRNA transport machinery in neurons, which move along microtubules. RNA granules contain densely packed...
journal_title:Cell structure and function
pub_type: 杂志文章,评审
doi:10.1247/csf.30.51
更新日期:2005-01-01 00:00:00
abstract::We have found that 1,9-Dideoxyforskolin (DDF) strongly inhibited the cell death induced by ricin, modeccin, Pseudomonas toxin, and diphtheria toxin in MDCK cells, suggesting that these protein toxins have a DDF-sensitive common pathway leading to cell death. However, no significant effect of forskolin on these toxins ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.22.545
更新日期:1997-10-01 00:00:00
abstract::Developing cultures of dissociated cerebral hemispheres obtained from 18-day-old embryonic rats synthesized, or activated, a myelin-related enzyme, 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase). This increase in activity coincided with the onset of myelination. The presence of CNPase in oligodendrocytes and mye...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.10.271
更新日期:1985-09-01 00:00:00
abstract::Recent studies have demonstrated that vesicle transport to cleavage furrow is indispensable for cytokinesis. Some animal and plant cells form distinct structures during cell division known as central spindle and phragmoplast, respectively. Several essential factors involved in the vesicle transport have been isolated ...
journal_title:Cell structure and function
pub_type: 杂志文章,评审
doi:10.1247/csf.26.567
更新日期:2001-12-01 00:00:00
abstract::When spores of Dictyostelium discoideum matured through normal development were stained with rhodamine-phalloidin, rod-shaped fluorescence was found across the nucleus and in the cytoplasm. Electron micrographs of the rod shape showed that it consisted of long and straight fibers 12 nm in diameter. In contrast, a conv...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.19.189
更新日期:1994-08-01 00:00:00
abstract::Synapsin I, one of the major synaptic proteins, is thought to associate with synaptic vesicles and to play a regulatory role in neurotransmitter release. In mature neurons, it is concentrated almost exclusively in presynaptic nerve endings. Here, we studied the subcellular localization of synapsin I during the develop...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.15.329
更新日期:1990-12-01 00:00:00
abstract::The Acanthamoeba myosin-IA and myosin-IB molecular motors bind to membranes, so they may produce the force to move organelles and membranes along actin filaments. We have determined the rate constants for the actin-activated myosin-I ATPase by pre-steady state kinetic analysis. ATP binds rapidly to myosin-I and dissoc...
journal_title:Cell structure and function
pub_type: 杂志文章,评审
doi:10.1247/csf.21.351
更新日期:1996-10-01 00:00:00
abstract::Stimulation of polymorphonuclear leukocytes (PMN) by tetravalent concanavalin A (alpha-ConA) induces membrane depolarization preceding the onset of superoxide anion (O2-) production. Both divalent and monovalent ConA analogues were studied to evaluate the role of valence. Monovalent ConA (m-ConA) was inactive in stimu...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.11.199
更新日期:1986-06-01 00:00:00
abstract::We have demonstrated simultaneous measurement of the membrane potential and cytoplasmic free Ca2+ concentration ([Ca2+]i) by utilizing a dual-laser flow cytometer in embryonic rat hippocampal cell suspensions. Veratrine, a Na+ channel activator, induced both membrane depolarization and elevation of [Ca2+]i. These acti...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.14.669
更新日期:1989-12-01 00:00:00
abstract::Cofilin, a 21kDa actin-binding protein, binds to F-actin in a 1:1 molar ratio of cofilin to actin molecule (Nishida, E., S. Maekawa, and H. Sakai, Biochemistry, 23, 5307-5313, 1984) and is capable of controlling actin polymerization and depolymerization in vitro in a pH-sensitive manner (Yonezawa, N., E. Nishida, and ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.12.443
更新日期:1987-10-01 00:00:00
abstract::We earlier discovered a novel 40-kDa protein (hsp40) induced by heat shock and other stresses in mammalian and avian cells. In this report, we purified the hsp40 in HeLa cells, using modified two-dimensional gel electrophoresis, and determined the amino terminal amino acid sequence of this protein. The hsp40 is homolo...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.17.77
更新日期:1992-02-01 00:00:00
abstract::Cultured fibroblasts from patients with Niemann-Pick disease type C (NP-C) are characterized by the lysosomal accumulation of unesterified cholesterol and the inability of low-density lipoprotein (LDL) to stimulate cholesterol esterification, in addition to impaired LDL-mediated down-regulation of LDL receptor activit...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.18.231
更新日期:1993-08-01 00:00:00
abstract::Newly prepared phosphatidylethanolamine (PE) conjugates of glycosaminoglycans (GAGs) enabled us to immobilize GAGs to solid phase through hydrophobic interaction. Using these compounds, called GAG-PEs, we studied the effects of GAGs immobilized to culture plates coated with various extracellular matrix (ECM) proteins ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.20.199
更新日期:1995-06-01 00:00:00
abstract::Analysis of the amino acid sequence in protein (deviation analysis) suggests that the binding between trypsin and the enveloped virus is the first step of their interaction, which occurs in a specified configuration. It is possible that the distance between their active sites is important for the viral sensitivity to ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.14.721
更新日期:1989-12-01 00:00:00
abstract::The purpose of the present study is to analyze the effect of serum or chick embryo extract (CEE) on the neuronal differentiation of the mouse neural crest cells. When the crest cells were cultured in the medium containing serum at low concentration (5% calf serum), neurite outgrowth was observed. The active outgrowth ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.13.267
更新日期:1988-06-01 00:00:00
abstract::A method has been developed to isolate pure preparations of nuclei in high yield from commercially available viable rice embryos (germ), employing extraction with buffer solution containing glycerol (without detergent) and polyamine, followed by centrifugation on a 30% Percoll cushion. The intactness of the isolated n...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.17.87
更新日期:1992-04-01 00:00:00