SYNTHESIS OF AN ENZYME DETERMINED BY AN ERYTHROCYTE NUCLEUS IN A HYBRID CELL

1969 ◽  
Vol 5 (1) ◽  
pp. 121-133
Author(s):  
H. HARRIS ◽  
P. R. HARRIS
Keyword(s):  
Mutant Mouse ◽  
Hybrid Cell ◽  
Critical Role ◽  
Surface Antigens ◽  
Mouse Cell ◽  
Inosinic Acid ◽  
Erythrocyte Nucleus ◽  
Transfer Of Information ◽  
Kinetics Of

When a chick erythrocyte nucleus is introduced into the cytoplasm of a mutant mouse cell lacking the enzyme inosinic acid pyrophosphorylase, synthesis of the enzyme is induced. However, this synthesis does not begin until the erythrocyte nucleus develops a nucleolus. The kinetics of synthesis of the enzyme are essentially similar to those previously described for the synthesis of surface antigens specified by the chick nucleus. The results of both sets of experiments indicate that the nucleolus plays a critical role in the transfer of information from the genes to the cytoplasm of the cell.

Species Specificity of an Enzyme Determined by an Erythrocyte Nucleus in an Interspecific Hybrid Cell

1970 ◽  
Vol 7 (1) ◽  
pp. 1-3
Author(s):  
P. R. COOK
Keyword(s):  
Interspecific Hybrid ◽  
Mutant Mouse ◽  
Hybrid Cell ◽  
Mouse Cell ◽  
Species Specificity ◽  
Inosinic Acid ◽  
Erythrocyte Nucleus

When a chick erythrocyte nucleus is introduced into the cytoplasm of a mutant mouse cell lacking inosinic acid pyrophosphorylase, synthesis of the enzyme is induced. The enzyme induced in this way has the characteristics of chick, not mouse, inosinic acid pyrophosphorylase.

The Expression of Genetic Information: A Study with Hybrid Animal Cells

1969 ◽  
Vol 4 (2) ◽  
pp. 499-525
Author(s):  
H. HARRIS ◽  
E. SIDEBOTTOM ◽  
D. M. GRACE ◽  
M. E. BRAMWELL
Keyword(s):  
Specific Surface ◽  
Genetic Information ◽  
Decisive Role ◽  
Surface Antigens ◽  
Mouse Cell ◽  
Animal Cells ◽  
Hybrid Animal ◽  
Specific Antigens ◽  
Erythrocyte Nucleus ◽  
Transfer Of Information

When the nucleus of a hen erythrocyte is introduced into the cytoplasm of a human or mouse cell in culture, it resumes the synthesis of RNA. The reactivated erythrocyte nucleus undergoes great enlargement, but it does not, for at least 2 or 3 days, develop nucleoli which can be discerned under the light microscope. During this period, the heterokaryon, although it may contain several active erythrocyte nuclei, does not synthesize any hen-specific surface antigens; and the hen-specific antigens introduced into the surface of the heterokaryon by the process of cell fusion are eliminated. But when, later, the erythrocyte nuclei do develop nucleoli, hen-specific antigens reappear on the surface of the heterokaryon and progressively accumulate. Before developing nucleoli, the erythrocyte nuclei synthesize little, if any, normal 28 S or 16 S RNA; but they do synthesize large amounts of the RNA which shows polydisperse sedimentation in conventional sucrose density gradients. Autoradiographic studies involving the use of a microbeam of ultraviolet light show, however, that this ‘polydisperse’ RNA is not transferred to the cytoplasm of the cell in detectable amounts so long as the erythrocyte nucleus lacks a definitive nucleolus. The inability of the erythrocyte nucleus at this stage to determine the synthesis of hen-specific surface antigens is thus attributable to the fact that it fails to transfer the RNA made on its chromosomes to the cytoplasm of the cell. When the erythrocyte nuclei develop nucleoli, however, the RNA which they make is transferred to the cytoplasm of the cell, and the synthesis of hen-specific surface antigens then begins. These experiments suggest that the nucleolus may play a decisive role in the transfer of information from nucleus to cytoplasm. The possible nature of this role is discussed.

scholarly journals QUANTITATIVE STUDIES ON THE MIXED LYMPHOCYTE INTERACTION IN RATS

1971 ◽  
Vol 134 (4) ◽  
pp. 857-870 ◽  
Author(s):  
Darcy B. Wilson ◽  
Dianne H. Fox
Keyword(s):  
Cell Surface ◽  
Surface Antigens ◽  
Mouse Cell ◽  
Cell Surface Antigens ◽  
Quantitative Studies ◽  
Lymphocyte Cultures ◽  
Mixed Lymphocyte Cultures ◽  
Environmental Antigens ◽  
Germfree Rats ◽  
Number Of Cells

The proliferative reactivity of lymphocytes from rat donors maintained under germfree or conventional conditions was examined in mixed lymphocyte cultures stimulated with allogeneic and xenogeneic cell surface antigens. The results show (a) that lymphocytes from conventionally maintained rats are less reactive to human, hamster, guinea pig, and mouse cell surface antigens than to the major H alloantigens, and (b) that lymphocytes from germfree rats display no demonstrable reactivity to xenogeneic cells, but are quantitatively normal in their response to allogenic cells. The conclusion drawn from these observations is that the circulating lymphocyte pool of an individual consists of a greater proportion of cells reactive to H alloantigens of other members of the same species than to the xenogeneic cellular antigens of members of other species and that this large number of cells is not generated by a mechanism involving immunization to cross-reactive environmental antigens.

Differentiation of two mouse cell lines is associated with hypomethylation of their genomes

1984 ◽  
Vol 4 (9) ◽  
pp. 1800-1806
Author(s):  
T H Bestor ◽  
S B Hellewell ◽  
V M Ingram
Keyword(s):  
Dna Methyltransferase ◽  
Cell Types ◽  
Mouse Cell ◽  
F9 Cells ◽  
Dna Restriction Fragments ◽  
Dna Restriction ◽  
F9 Embryonal Carcinoma Cells ◽  
Murine Erythroleukemia ◽  
Kinetics Of

Methyl-accepting assays and a sensitive method for labeling specific CpG sites have been used to show that the DNA of F9 embryonal carcinoma cells decreases in 5-methylcytosine content by ca. 9% during retinoic acid-induced differentiation, whereas the DNA of dimethyl sulfoxide-induced Friend murine erythroleukemia (MEL) cells loses ca. 3.8% of its methyl groups. These values correspond to the demethylation of 2.2 X 10(6) and 0.9 X 10(6) 5'-CpG-3' sites per haploid genome in differentiating F9 and MEL cells, respectively. Fluorography of DNA restriction fragments methylated in vitro and displayed on agarose gels showed that demethylation occurred throughout the genome. In uninduced F9 cells, the sequence TCGA tended to be more heavily methylated than did the sequence CCGG, whereas this tendency was reversed in MEL cells. The kinetics of in vitro DNA methylation reactions catalyzed by MEL cell DNA methyltransferase showed that substantial numbers of hemimethylated sites accumulate in the DNA of terminally differentiating F9 and MEL cells, implying that a partial loss of DNA-methylating activity may accompany terminal differentiation in these two cell types.

scholarly journals The 27-kD diphtheria toxin receptor-associated protein (DRAP27) from vero cells is the monkey homologue of human CD9 antigen: expression of DRAP27 elevates the number of diphtheria toxin receptors on toxin-sensitive cells.

1992 ◽  
Vol 118 (6) ◽  
pp. 1389-1399 ◽  
Author(s):  
T Mitamura ◽  
R Iwamoto ◽  
T Umata ◽  
T Yomo ◽  
I Urabe ◽  
...  
Keyword(s):  
Diphtheria Toxin ◽  
Binding Capacity ◽  
Hybrid Cell ◽  
Antigen Expression ◽  
Vero Cells ◽  
Mouse Cell ◽  
Homology Search ◽  
Receptor Associated Protein ◽  
New Family ◽  
Cloned Cdna

Diphtheria toxin (DT) receptor associates with a 27-kD membrane protein (DRAP27) in monkey Vero cells. A cDNA encoding DRAP27 was isolated, and its nucleotide sequence was determined. The deduced amino acid sequence revealed that DRAP27 is the monkey homologue of human CD9 antigen. DRAP27 is recognized by CD9 antibodies. A human-mouse hybrid cell line (3279-10) possessing human chromosome 5, sensitive to DT, but not expressing CD9 antigen, was used for transfection experiments with DRAP27. When the cloned cDNA encoding DRAP27 was transiently expressed in 3279-10 cells, the total DT binding capacity was three to four times higher than that of untransfected controls. Transfectants stably expressing DRAP27 have an increased number of DT binding sites on the cell surface. Furthermore, the transfectants are 3-25 times more sensitive to DT than untransfected cells, and the sensitivity of these cells to DT is correlated with the number of DRAP27 molecules on the surface. However, when the cloned cDNA was introduced into mouse cell lines that do not express DT receptors, neither an increased DT binding nor enhancement of DT sensitivity was observed. Hence, we conclude that DRAP27 itself does not bind DT, but serves to increase DT binding and consequently enhances DT sensitivity of cells that have DT receptors. 12 proteins related to DRAP27/CD9 antigen were found through homology search analysis. These proteins appear to belong to a new family of transmembrane proteins.

scholarly journals Batch Syngas Fermentation by Clostridium carboxidivorans for Production of Acids and Alcohols

Processes ◽  
2020 ◽  
Vol 8 (9) ◽  
pp. 1075
Author(s):  
Fabiana Lanzillo ◽  
Giacomo Ruggiero ◽  
Francesca Raganati ◽  
Maria Elena Russo ◽  
Antonio Marzocchella
Keyword(s):  
Critical Role ◽  
Volume Ratio ◽  
Liquid Volume ◽  
Syngas Fermentation ◽  
Co Concentration ◽  
Depletion Rate ◽  
Growth Differences ◽  
Gas Volume ◽  
Clostridium Carboxidivorans ◽  
Kinetics Of

Syngas (CO, CO2, and H2) has attracted special attention due to the double benefit of syngas fermentation for carbon sequestration (pollution reduction), while generating energy. Syngas can be either produced by gasification of biomasses or as a by-product of industrial processes. Only few microorganisms, mainly clostridia, were identified as capable of using syngas as a substrate to produce medium chain acids, or alcohols (such as butyric acid, butanol, hexanoic acid, and hexanol). Since CO plays a critical role in the availability of reducing equivalents and carbon conversion, this work assessed the effects of constant CO partial pressure (PCO), ranging from 0.5 to 2.5 atm, on cell growth, acid production, and solvent production, using Clostridium carboxidivorans. Moreover, this work focused on the effect of the liquid to gas volume ratio (VL/VG) on fermentation performances; in particular, two VL/VG were considered (0.28 and 0.92). The main results included—(a) PCO affected the growth kinetics of the microorganism; indeed, C. carboxidivorans growth rate was characterized by CO inhibition within the investigated range of CO concentration, and the optimal PCO was 1.1 atm (corresponding to a dissolved CO concentration of about 25 mg/L) for both VL/VG used; (b) growth differences were observed when the gas-to-liquid volume ratio changed; mass transport phenomena did not control the CO uptake for VL/VG = 0.28; on the contrary, the experimental CO depletion rate was about equal to the transport rate in the case of VL/VG = 0.92.

Correction of Genetic Defects in Mammalian Cells by the Input of Small Amounts of Foreign Genetic Material

1973 ◽  
Vol 13 (3) ◽  
pp. 841-861
Author(s):  
YVONNE L. BOYD ◽  
H. HARRIS
Keyword(s):  
Thymidine Kinase ◽  
Mammalian Cells ◽  
Hybrid Cell ◽  
Genetic Material ◽  
Chinese Hamster ◽  
Heat Sensitivity ◽  
Chinese Hamster Cells ◽  
Inosinic Acid ◽  
Mouse Cells

Chinese hamster cells lacking inosinic acid pyrophosphorylase and mouse cells lacking thymidine kinase were fused with chick erythrocytes. The resultant heterokaryons were cultivated in a selective medium in which possession of these enzymes was essential for cell survival and growth. Clones of cells able to grow in this medium were isolated and studied. A detailed karyological analysis of these clones failed to reveal any chick chromosomes; nor could any chick-specific antigens be detected on the surface of the cells. Nonetheless, clones arising from the fusion of chick erythrocytes with Chinese hamster cells were shown to possess an inosinic acid pyrophosphorylase which had the electrophoretic characteristics of chick inosinic acid pyrophosphorylase. However, the clones arising from the fusion of the chick erythrocytes with the mouse cells had a thymidine kinase with the electrophoretic mobility and heat sensitivity of murine, not chick, thymidine kinase. Both types of hybrid cell have now been cultivated in vitro for 18 months without the loss of thymidine kinase or inosinic acid pyrophosphorylase activity.

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