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41.
Summary We have isolated Saccharomyces cerevisiae mutants, smp, showing stable maintenance of plasmid pSRI, a Zygosaccharomyces rouxii plasmid. The smp mutants were recessive and were classified into at least three different complementation groups. The three mutants also showed increased stability of YRp plasmids and the mutations are additive for plasmid stability. One mutation, smp1, confers a respiration-deficient (rho
0) phenotype and several Rho– mutants independently isolated by ethidium bromide treatment of the same yeast strain also showed increased stabilities of pSR1 and YRp plasmids. The wild-type S. cerevisiae cells showed a strongly biased distribution of pSR1 molecules as well as YRp plasmids to the mother cells at mitosis, while the smpf mutant did not show this bias. Another mutation, smp3, at a locus linked to ade2 on chromosome XV, confers temperature-sensitive growth. The SMP3 gene encodes a 59.9 kDa hydrophobic protein and disruption of the gene is lethal. 相似文献
42.
Kazuhiro Yoshikawa Masao Seto Ryuzo Ueda Yuichi Obata Kunihiro Notake Takashi Yokochi Toshitada Takahashi 《Immunogenetics》1991,33(5-6):352-360
The CD7 molecule is a differentiation antigen found on the surface of T lymphocytes and also on a very minor fraction of acute nonlymphocytic leukemia (ANLL). To study the genomic structure of the CD7 gene, two clones (SY4 and SY22) were isolated by screening a genomic library with a CD7 cDNA probe. Restriction mapping of these two phage clones showed that both overlapped each other, covering a total length of 23 kilobases (kb). Transfection of mouse L cells demonstrated that SY22 contains the gene expressing the CD7 antigen reactive with monoclonal CD7 antibody (Tp40), while SY4 does not. Subcloning of a 10.5 kb fragment from a 14.4 kb insert of SY22 contained the structural gene for the CD7 antigen. Detailed restriction mapping and partial sequence analysis revealed the CD7 gene to consist of four exons. By RNase protection assay, multiple initiation sites — 122 base pairs (bp) to — 38 bp from ATG translation initiation site were demonstrated. The promoter region had high G+C content and contained two SP1 binding sites (CCGCCC) and an AP2 binding site (CCCCAGGC), but lacked CAAT and TATA motifs. 相似文献
43.
Summary The isomerization of D-glucose in mixed ethanol-water was studied at various reaction temperatures (40–70 °C), employing glucose isomerase fromStreptomyces phaeochromogenes andClostridium thermohydrosulfuricum, respectively. The thermophilicClostridium enzyme was considerably, more stable towards the combination of organic cosolvent and increased temperature and with this enzyme a 55% yield of fructose from glucose was obtained at relatively low concentration of ethanol (40 %). 相似文献
44.
Conformational dynamics of DNA affected by intercalation and minor groove binding: direct observation of large DNA. 总被引:1,自引:0,他引:1
Y Matsuzawa K Minagawa K Yoshikawa M Matsumoto M Doi 《Nucleic acids symposium series》1991,(25):131-132
Using fluorescence microscopy, we have observed moving DNA molecules in solution and analyzed the "higher-order" structure in a quantitative manner. It was found that EB (ethidium bromide), an intercalator, has the effect to increase the persistent length. In other words, EB expands DNA. Whereas, DAPI (4',6-diamidino-2-phenylindole), a minor groove binding drug, decreases the persistent length. It is demonstrated that the direct observation of DNA molecules with fluorescence microscopy is quite useful to study the interaction of various chemical compounds with DNA molecules. 相似文献
45.
46.
Kenji Tomioka Kenji Yamada Shinya Yokoyama Yoshihiko Chiba 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1991,169(3):291-298
The coupling mechanism between the bilaterally paired optic lobe circadian pacemakers in the cricket Gryllus bimaculatus was investigated by recording locomotor activity, under constant light or constant red light, after the optic nerve was unilaterally severed.
These results suggest that the 2 optic lobe pacemakers weakly couple to one another and that the cricket maintains a stable temporal structure in its behavior through the phase-dependent mututal inhibition of activity and the phase-dependent freerunning period modulation. 相似文献
1. | The majority (about 70%) of the animals showed a locomotor rhythm with 2 rhythmic components; one freerunning with a period of 25.33 ± 0.41 (SD) h and the other with 24.36 ± 0.37 (SD) h under constant light (Fig. 3A). |
2. | Removal of the intact side optic lobe abolished the longer period component (Fig. 4A), while the operation on the operated side caused a reverse effect (Fig. 4B), indicating that the longer and the shorter period components are driven by the pacemaker on the intact and the operated side, respectively. |
3. | The activity driven by a pacemaker was inhibited during the subjective day of the contralateral pacemaker (circadian time 0–10, Fig. 5). |
4. | The freerunning periods of the two components were not constant but varied as a function of the mutual phase angle relationship (Figs. 3A, 7, 8). |
47.
A potential approach for gene therapy targeting hepatoma using a liver-specific promoter on a retroviral vector. 总被引:7,自引:0,他引:7
S Kuriyama M Yoshikawa S Ishizaka T Tsujii K Ikenaka T Kagawa N Morita K Mikoshiba 《Cell structure and function》1991,16(6):503-510
Recent technological advances made in molecular biology and in vitro culture of human and other mammalian cells have led to broad medical and scientific acceptance of the feasibility of gene therapy for genetic diseases. Cancer might practically be one of the attractive targets for such therapy. For the treatment of cancer, it is important to manipulate the gene of interest such that it is expressed solely in cancer cells. We have developed a tissue-specific gene expression system, based on a tissue-specific promoter on a retroviral vector. A murine ecotropic retroviral vector was constructed in which the Escherichia coli beta-galactosidase gene served as a reporter; it was expressed under control of the albumin enhancer element and promoter. The tissue specificity of this vector was first assessed in vitro, and beta-galactosidase activity was detected exclusively in hepatoma cell lines. This recombinant retrovirus was injected directly into a subcutaneous tumor composed of transplantable murine MH-134 hepatoma cells, and expression of the gene was observed in vivo. Then this recombinant retrovirus was injected via the spleen or directly into the liver, resulting in the gene expression in dividing hepatocytes in partially hepatectomized mice, but not in nondividing hepatocytes in normal mice. Gene transfer specific to dividing hepatocytes and expression by means of retroviral vectors should possess high potential for selective elimination of hepatoma cells surrounded by nondividing normal hepatocytes. 相似文献
48.
Expression of the enkephalin precursor gene in rat Sertoli cells. Regulation by follicle-stimulating hormone 总被引:1,自引:0,他引:1
Searching for somatic cells expressing the preproenkephalin (A) gene in the testis, we have isolated Sertoli cells from the testes of 20-day-old rats. Cultured Sertoli cells contained a single species (about 1.5 kb) of preproenkephalin mRNA, and follicle-stimulating hormone (FSH) transiently increased the mRNA abundance to a maximum (about 30 molecules per cell) at 12 h. Various compounds that activate the cyclic AMP system in Sertoli cells similarly increased the abundance of preproenkephalin mRNA. Moreover, FSH increased intracellular Met-enkephalin immunoreactive peptides in Sertoli cells. Thus, the preproenkephalin gene expression in Sertoli cells is positively regulated by FSH through the cyclic AMP system. 相似文献
49.
Interaction of the Bacillus subtilis DnaA-like protein with the Escherichia coli DnaA protein. 总被引:5,自引:2,他引:3 下载免费PDF全文
Plasmids carrying the intact Bacillus subtilis dnaA-like gene and two reciprocal hybrids between the B. subtilis and Escherichia coli dnaA genes were constructed. None of the plasmids could transform wild-type E. coli cells unless the cells contained surplus E. coli DnaA protein (DnaAEc). A dnaA (Ts) strain integratively suppressed by the plasmid R1 origin could be transformed by plasmids carrying either the B. subtilis gene (dnaABs) or a hybrid gene containing the amino terminus of the E. coli gene and the carboxyl terminus of the B. subtilis gene (dnaAEc/Bs). In cells with surplus E. coli DnaA protein, expression of the E. coli dnaA gene was derepressed by the B. subtilis DnaA protein and by the hybrid DnaAEc/Bs protein, whereas it was strongly repressed by the reciprocal hybrid protein DnaABs/Ec. The plasmids carrying the different dnaA genes probably all interfere with initiation of chromosome replication in E. coli by decreasing the E. coli DnaA protein concentration to a limiting level. The DnaABs and the DnaAEc/Bs proteins effect this decrease possibly by forming inactive oligomeric proteins, while the DnaABs/Ec protein may decrease dnaAEc gene expression. 相似文献
50.
Virulence-associated genetic regions comprising 31 kilobases of the 230-kilobase plasmid in Shigella flexneri 2a. 总被引:42,自引:9,他引:33 下载免费PDF全文
C Sasakawa K Kamata T Sakai S Makino M Yamada N Okada M Yoshikawa 《Journal of bacteriology》1988,170(6):2480-2484
By random transposon Tn5 insertions, we previously identified six virulence-associated SalI fragments, B, D, F, G, H, and P, in the 230-kilobase plasmid pMYSH6000 of Shigella flexneri 2a. In this study, we analyzed the sites of 134 independent Tn5 insertions on four contiguous SalI fragments, B, P, H, and D, of pMYSH6000 and identified five virulence-associated regions; four were associated with inducing a positive Sereny test (Ser), invasion into epithelial cells (Inv), binding to Congo red (Pcr), and inhibition of bacterial growth (Igr), and one was associated with the Ser and Inv but not with the Pcr or Igr phenotypes. Hybridization studies revealed that these virulence-associated DNA regions were highly conserved among 15 other virulence plasmids of four species of Shigella and enteroinvasive Escherichia coli. These data indicate that at least seven separate genetic determinants on the virulence plasmid are required for full expression of the virulence phenotype of shigellae. 相似文献