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91.
92.
Sumitomo Shinichiro Tatemoto Yukihiro Fukui Shin Nakamura Taka-aki Fukushima Shoji Ito Nobuyuki Mori Masahiko 《Virchows Archiv. B, Cell pathology including molecular pathology》1985,49(1):395-399
Virchows Archiv B Cell Pathology - Paget cells from cases of mammary and extramammary Paget’s disease were examined for carcinoembryonic antigen (CEA) and CEA-related antigens by the... 相似文献
93.
Isolation and physical mapping of temperature-sensitive mutants defective in heat-shock induction of proteins in Escherichia coli 总被引:25,自引:0,他引:25
Summary Mutants of Escherichia coli K12 that are partially or totally defective in induction of major heat-shock proteins and cannot grow at high temperature (42° C) were isolated by localized mutagenesis. These mutants carry a single mutation in the gene htpR (formerly hin) located at min 76 on the E. coli genetic map. Some mutants exhibit delayed (partial) induction of heat-shock proteins or require a higher temperature for induction than the wild type, whereas others are not induced under any of these conditions. The maximum temperature that allows growth varies among different mutants and is correlated with the residual induction capacity. Temperature-resistant revertants obtained from each mutant are fully or partially recovered in heat-shock induction. These results indicate that the inability of htpR mutants to grow at high temperature is due to the defect in heat-shock induction. In addition, a couple of mutants was found that produce significantly higher amounts of heat-shock proteins even at 30° C.The htpR gene has been cloned into plasmid pBR322 using the above mutants, and was localized to a DNA segment of 1.6 kilobase pairs. The mutants harboring certain palsmids that carry a part of htpR produce temperature-resistant recombinants at high frequency. This permits further localization of mutations within the htpR gene. Analysis of proteins encoded by each of the recombinant plasmids including the one carrying a previously isolated amber mutation (htpR165) led to the identification of a protein with an apparent molecular weight of about 36,000 daltons as the htpR gene product. 相似文献
94.
Identification of the secY (prlA) gene product involved in protein export in Escherichia coli 总被引:17,自引:0,他引:17
Koreaki Ito 《Molecular & general genetics : MGG》1984,197(2):204-208
Summary The gene secY (or prlA) is essential for protein export across the cytoplasmic membrane of Escherichia coli. The protein product of secY has been identified using the gene cloned under the control of the pL promoter of phage in combination with the maxicell system. The protein was found to have some unusual properties. First, it is important not to heat the protein at 100°C in the SDS sample buffer for its subsequent detection by gel electrophoresis. Second, migration of the protein in SDS-polyacrylamide gel electrophoresis is variable depending on the gel compositions. Gels with stronger sieving effect give higher apparent molecular weights. These properties are similar to those of hydrophobic proteins of the cytoplasmic membrane, such as the lactose permease. Finally, a major fraction of the protein synthesized from the overproducing plasmid is degraded rapidly in vivo. The altered protein from the secY24 mutant gene is even more unstable. These results provide information which is basic for the dissection of the protein export machinery of the bacterial cell. 相似文献
95.
Enrichment and characterization of clonogenic epithelial cells from adult rat liver and initiation of epithelial cell strains 总被引:8,自引:0,他引:8
Kazunori Furukawa Tomiko Shimada Patricia England Yohichi Mochizuki Gary M. Williams 《In vitro cellular & developmental biology. Plant》1987,23(5):339-348
Summary A highly efficient method is described for obtaining prolifertive epithelial cells from adult rat livers for the reproducible
establishment of liver epithelial cell strains. When cells were isolated from livers of 10-to 15-wk-old male Fischer 344 rats
by a collagenase-perfusion method, collected by centrifugation at 50×g for 5 min, and cultured in Williams' medium E containing fetal bovine serum and dexamethasone, colonies of epithelial cells
different in size and morphology from hepatocytes were obtained. Sequential perfusion with collagenase and dispase yielded
numerous epithelial cell colonies. When isolated cells were fractionated by differential centrifugation, the great majority
of hepatocytes were sedimented at 50 ×g for 1 min, whereas many non-hepatocytic cells remiined in the supernatant and could be sedimented by a second centrifugation
at 50×g for 5 min. Culture of the two fractions revealed that almost all the epithelial cell colonies were derived from cells in
the non-hepatocytic cell fraction. The epithelial cells were cytochemically negative for γ-glutamyl transpeptidase activity,
whereas an increase in the activity was detected in hepatocytes with duration in culture. Ultrastructural characteristics
of hepatocytes were not found in the cells of newly established cell strains. These results suggest that adult rat liver epithelial
cells propagable in culture were derived from a cell type other than the hepatocyte. 相似文献
96.
Hideya Hayashi Seiji Ito Teruo Tanaka Manabu Negishi Hideo Kawabe Yokohama Hiromitsu Kikuko Watanabe Osamu Hayaishi 《Prostaglandins & other lipid mediators》1987,33(4)
In view of the recent finding that prostaglandin D2 is stereospecifically converted to 9α,11β-prostaglandin F2, an isomer of prostaglandin F2α, a highly specific and sensitive radioimmunoassay for 9α,11β-prostaglandin F2 was developed and applied to determine the content of this prostaglandin in various rat tissues. Antisera against 9α-11β-prostaglandin F2 were raised in rabbits immunized with the bovine serum albumin conjugate, and [3H]9α,11β-prostaglandin F2 was enzymatically prepared from [3H]prostaglandin D2. The assay detected 9α,11β-prostaglandin F2 over the range of 20 pg to 1 ng, and the antiserum showed less than 0.04% cross-section with prostaglandin F2α, prostaglandin F2β and 9β,11β-prostaglandin F2. To avoid postmortem changes, tissues were frozen in liquid nitrogen immediately after removal. The basal level of 9α,11β-prostaglandin F2 was hardly detectable in various tissues of the rat examined, including spleen, lung, liver and brain; although it was found to be 0.31 ± 0.06 ng/g wet weight in the small intestine. During convulsion induced by pentylenetetrazole, enormous amounts of prostaglandin D2 (ca. 180 ng/g wet weight) and prostaglandin F2α (ca. 70 ng/g) were produced in the brain; however, 9α,11β-prostaglandin F2 was detected neither there nor in the blood. This result demonstrates that the conversion to 9α,11β-prostaglandin F2 is a minor pathway, if one at all, of prostaglandin D2 metabolism in the rat brain. 相似文献
97.
98.
99.
Altered translation initiation factor 2 in the cold-sensitive ssyG mutant affects protein export in Escherichia coli. 总被引:10,自引:0,他引:10 下载免费PDF全文
The Escherichia coli gene secY (pr1A) codes for an integral membrane protein that plays an essential role in protein export. We previously isolated cold-sensitive mutations (ssy) as extragenic suppressors of temperature-sensitive secY24 mutation. Now we show that the ssyG class of mutations are within infB coding for the translation initiation factor IF2. The mutants produce altered forms of IF2 with a cold-sensitive in vitro activity to form a translation initiation complex. The mutation suppresses not only secY24 but also other secretion-defective mutations such as secA51 and rp10215. The beta-galactosidase enzyme activity of the MalE-LacZ 72-47 hybrid protein is strikingly reduced in the ssyG mutant at the permissive high temperature, while the hybrid protein itself is normally synthesized. This effect, which was observed only for the hybrid protein with a functional signal sequence, may result from some alteration in the cellular localization of the protein. These results suggest that IF2 or the translation initiation step can modulate protein export reactions. The isolation of cold-sensitive ssyG mutations in infB provides genetic evidence that IF2 is indeed essential for normal growth of E. coli cells. 相似文献
100.
Nucleotide sequence of Bacillus phage phi 29 genes 14 and 15: homology of gene 15 with other phage lysozymes. 总被引:8,自引:0,他引:8 下载免费PDF全文
The nucleotide sequence of Bacillus phage phi 29 genes 14 (g14) and 15 (g15) have been determined and shown to encode proteins with molecular weights of 15,014 and 28,022, respectively. The g14 open reading frame (ORF) was confirmed by sequencing a sus14(1241) mutant. Gene product 15 (gp15) has considerable homology with Salmonella phage P22 lysozyme and lesser homology with Escherichia coli phage T4 lysozyme. Putative translation signals are identified. In addition, the role of a previously described promoter, B2, is discussed. 相似文献