Author Oda, Takuzo| Seki, Shuji| Watanabe, Sekiko|
Published Date 1969-10
Publication Title Acta Medicinae Okayama
Volume volume23
Issue issue5
Content Type Journal Article
JaLCDOI 10.18926/AMO/32400
FullText URL fulltext.pdf
Author Oda, Takuzo| Watanabe, Sekiko| Nakamura, Takashi|
Abstract <p>Electron microscopy of four human T-cell lines revealed the production of type C virus particles in two T-cell lines: one derived from acute lymphoblastic leukemia and the other from a leukemic T-lymphoid malignancy. Virus particles isolated from these cells had reverse transcriptase activity and the major internal structural protein of 30,000 daltons (p30). The indirect immunofluorescence test of these virus-producing cells with sera of patients with adult T-cell leukemia (ATL) was negative. The data indicate that these retroviruses are different from adult T-cell leukemia virus (ATLV).</p>
Keywords type C virus particles human T-cell lines electron microscopy virion proteins immunofluorescence test
Amo Type Article
Published Date 1983-12
Publication Title Acta Medica Okayama
Volume volume37
Issue issue6
Publisher Okayama University Medical School
Start Page 529
End Page 533
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 6198871
Web of Sience KeyUT A1983RW62800011
JaLCDOI 10.18926/AMO/32188
FullText URL fulltext.pdf
Author Zhang, Bo| Watanabe, Sekiko| Akiyama, Kosuke| Li, Ting| Fukushima, Keisuke| Tsutsui, Ken| Seki, Shuji|
Abstract <p>DNA repair synthesis induced in permeable mouse ascites sarcoma cells by peplomycin, an antitumor antibiotic, was studied. Mouse ascites sarcoma (SR-C3H/He) cells were permeabilized with a low concentration of Triton X-100 in an isotonic condition. Permeable cells were treated with an appropriate concentration of peplomycin to introduce single-strand breaks in permeable cell DNA. DNA repair synthesis in peplomycin-treated permeable cells was measured by incubating the cells with four deoxynucleoside triphosphates in an appropriate buffer system. The DNA repair synthesis was enhanced by ATP and NaCl at near physiological concentrations. More than 90% of DNA synthesis in the present system depended on the peplomycin-treatment. The repair nature of the DNA synthesis was confirmed by a BrdUMP density shift technique. The repair patches were largely completed and ligated in the presence of ATP. Analyses using selective inhibitors for DNA polymerases showed that both DNA polymerase Beta and aphidicolin-sensitive DNA polymerases (DNA polymerase alpha and/or delta) were involved in the repair DNA synthesis.&#60;/P&#62;</p>
Keywords DNA repair peplomycin DNA polymerases permeable mouse cells
Amo Type Article
Published Date 1991-04
Publication Title Acta Medica Okayama
Volume volume45
Issue issue2
Publisher Okayama University Medical School
Start Page 89
End Page 94
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 1714230
Web of Sience KeyUT A1991FL60800004
JaLCDOI 10.18926/AMO/31276
FullText URL fulltext.pdf
Author Ocho, Mumehiko| Nakai, Satoru| Tasaka, Kenji| Watanabe, Sekiko| Oda, Takuzo|
Abstract <p>Simian virus 40 (SV40) DNA was microinjected into cultured mammalian cells by means of electrophoresis (iontophoresis). Successful transfer of DNA into cells was confirmed by detecting SV40 T antigen using the indirect immunofluorescent technique.</p>
Keywords microinjection electrophoresis SV40 DNA
Amo Type Brief Note
Published Date 1981-11
Publication Title Acta Medica Okayama
Volume volume35
Issue issue5
Publisher Okayama University Medical School
Start Page 381
End Page 384
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 6274168
Web of Sience KeyUT A1981MS42400009
JaLCDOI 10.18926/AMO/31271
FullText URL fulltext.pdf
Author Oda, Takuzo| Watanabe, Sekiko| Hanakawa, Shiro| Hosogi, Nobuo|
Abstract <p>A permeable cell system has been developed by treatment with saponin for studying in vitro replication of DNA and chromatin. DNA replication of simian virus 40 nucleoprotein complexes (SV40 chromatin) in saponin-treated permeable cells was found to be more efficient than that in digitonin-treated permeable cells. Autoradiography of the agarose-gel revealed that [alpha-32P]dCTP was incorporated into SV40 DNA I, II and replicating intermediates. The time course of the incorporation indicated complete replication of SV40 DNA and chromatin with a full number of nucleosomes. The saponin-treated permeable cell system will serve as a useful system for studying in vitro replication of DNA and chromatin in eukaryotic cells.</p>
Keywords saponin permeable cells DNA replication in vitro SV40 Chromatin replication gel-electrophoresis autoradiography.
Amo Type Brief Note
Published Date 1981-04
Publication Title Acta Medica Okayama
Volume volume35
Issue issue2
Publisher Okayama University Medical School
Start Page 149
End Page 154
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 6269361
Web of Sience KeyUT A1981LS45700008
Author Tanabe, Naoko| Hidaka, Hideyuki| Watanabe, Sekiko| Oda, Takuzo|
Published Date 1978-12
Publication Title Acta Medica Okayama
Volume volume32
Issue issue6
Content Type Journal Article
JaLCDOI 10.18926/AMO/30869
FullText URL fulltext.pdf
Author Ikeda, Shogo| Tsutsui, Ken| Hatsushika, Masao| Watanabe, Sekiko| Oda, Takuzo|
Abstract <p>The major gag protein (p34) of squirrel monkey retrovirus-H was purified in one chromatographic step by anion-exchange high performance liquid chromatography. The virus in a crude fraction was disrupted with Brij 35 in the presence of three kinds of protease inhibitors. The soluble virus lysate was injected into a Polyanion SI column, and p34 was eluted with a linear salt gradient. The recovery of the protein was about 60%. The purified p34 was nearly homogenous as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining.</p>
Keywords retrovirus gag protein protein purification high performance liquid chromatography
Amo Type Article
Published Date 1989-04
Publication Title Acta Medica Okayama
Volume volume43
Issue issue2
Publisher Okayama University Medical School
Start Page 127
End Page 129
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 2786318
Web of Sience KeyUT A1989U578500007
JaLCDOI 10.18926/AMO/30857
FullText URL fulltext.pdf
Author Ikeda, Shogo| Yamamoto, Mihoko| Nagao, Kazutaka| Zhang, Bo| Watanabe, Sekiko| Oda, Takuzo|
Abstract <p>Non-radioactive hybridization probes were prepared using the M13 phage vector and the universal sequencing primer. The probe sequence to be used was first cloned into the M13 vector, and the minus strand of the template DNA was then synthesized with the Klenow fragment of E. coli DNA polymerase I in the presence of the biotinylated nucleotide, biotin-11-dUTP, as a label. Resultant DNA was heavily biotinylated, and made up of the entire minus strand of the template DNA. The long tag sequence derived from the M13 vector may increase the sensitivity of the detection. The biotinylated hybrids were visualized with the streptavidin-alkaline phosphatase conjugate and chromogenic substrates. As shown by Southern hybridization, the probe prepared in this way could be used to detect less than 1 pg of target sequence and a single copy gene sequence in human genomic DNA within several hours of signal development.</p>
Keywords non-rradioctive probe biotin nucleotide M13 phage DNA universal sequencing primer Southern hybridization
Amo Type Article
Published Date 1989-08
Publication Title Acta Medica Okayama
Volume volume43
Issue issue4
Publisher Okayama University Medical School
Start Page 197
End Page 202
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 2678902
Web of Sience KeyUT A1989AP79100001
Author Oda, Takuzo| Watanabe, Sekiko| Nakamura, Takashi| Hidaka, Hideyuki|
Published Date 1977-12
Publication Title Acta Medica Okayama
Volume volume31
Issue issue6
Content Type Journal Article
JaLCDOI 10.18926/AMO/30763
FullText URL fulltext.pdf
Author Nakagawa, Yuko| Watanabe, Sekiko| Akiyama, Kosuke| Sarker, Altaf H| Tsutsui, Ken| Inoue, Hajime| Seki, Shuji|
Abstract <p>We purified a 44-kDa nuclear protein from salt-extract of permeable mouse ascites sarcoma cells in an effort to isolate factors involved in the repair of acid-depurinated DNA. It was copurified with a major AP endonuclease (APEX nuclease) by sequential column chromatography then further purified by sodium dodecyl sulphate-poly-acrylamide gel electrophoresis as a possible DNA repair support factor. Its partial amino acid sequences were determined, and a cDNA clone for the protein was isolated from a mouse T-cell cDNA library using long degenerate oligonucleotide probes deduced from the amino acid sequence. The complete nucleotide sequence of the cDNA (1.7 kilobases) was determined. Northern hybridization using this cDNA detected two transcripts: 1.8kb being the major one and 2.6 kb being the minor one. The complete amino acid sequence for the protein predicted from the nucleotide sequence of the cDNA indicates that the 44-kDa protein consists of 394 amino acids with a calculated molecular weight of 43,698. In tests performed thus far, the recombinant 44-kDa protein expressed in Escherichia coli has not expressed any repair-support activity. It remains to be analyzed whether the protein attains this activity after appropriate posttranslational modifications. Most parts of the 44-kDa protein cDNA and the deduced amino acid sequence were found to be identical to those of the protein p38 -2G4, recently reported as a cell cycle-specifically modulated nuclear protein of 38kDa. The p38-2G4 may be a truncated form of the present 44-kDa protein.</p>
Keywords 44-kDa protein nuclear protein cDNA cloning cDNA sequencing recombinant protein
Amo Type Article
Published Date 1997-08
Publication Title Acta Medica Okayama
Volume volume51
Issue issue4
Publisher Okayama University Medical School
Start Page 195
End Page 206
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 9284967
Web of Sience KeyUT A1997XU03200003
JaLCDOI 10.18926/AMO/30527
FullText URL fulltext.pdf
Author Oda, Takuzo| Watanabe, Sekiko| Hanakawa, Shiro| Nakamura, Takashi|
Abstract <p>A permeable cell system has been developed by treatment with digitonin for studying in vitro DNA replication of chromatin. DNA replication of simian virus 40 nucleoprotein complexes (SV40 chromatin) in digitonin-treated permeable cells was analyzed by electrophoresis in agarose-gel. Autoradiography of the agarose-gel revealed that [32P]dCTP was incorporated in SV40 DNA I, II and replicating intermediates. The time course of the incorporation indicated the complete replication of SV40 DNA and chromatin with a full number of nucleosomes. The digitonin-treated permeable cell system will serve as a useful system for studying in vitro DNA replication of chromatin.</p>
Keywords digitonin permeable cells DNA replication in vitro SV40 chromatin replication gel -electrophoresis autoradiography
Amo Type Brief Note
Published Date 1980-12
Publication Title Acta Medica Okayama
Volume volume34
Issue issue6
Publisher Okayama University Medical School
Start Page 409
End Page 413
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 6258398
Web of Sience KeyUT A1980KZ17800007
JaLCDOI 10.18926/AMO/30311
FullText URL fulltext.pdf
Author Ikeda, Shogo| Hatsushika, Masao| Shigehara, Tsuguya| Watanabe, Sekiko| Omura, Sachiko| Tsutsui, Ken| Oda, Takuzo|
Abstract <p>Simian virus 40 (SV40) large T antigen was partially purified from small amounts of SV40-infected and SV40-transformed cells by immunoaffinity chromatography with high recovery. T antigen, in both crude and partially purified states, was detected rapidly by a sensitive and quantitative enzyme-linked immunosorbent assay (ELISA). Stability of the partially purified T antigen was found to increase by addition of 0.01% bovine serum albumin (BSA).</p>
Keywords SV40 T antigen affinity chromatography ELISA
Amo Type Article
Published Date 1984-08
Publication Title Acta Medica Okayama
Volume volume38
Issue issue4
Publisher Okayama University Medical School
Start Page 341
End Page 347
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 6093443
Web of Sience KeyUT A1984TG25900003