scholarly journals Isolation of microcell hybrid clones containing retroviral vector insertions into specific human chromosomes.

1987 ◽  
Vol 7 (8) ◽  
pp. 2814-2820 ◽  
Author(s):  
T G Lugo ◽  
B Handelin ◽  
A M Killary ◽  
D E Housman ◽  
R E Fournier
Keyword(s):  
High Efficiency ◽  
Human Fibroblasts ◽  
Retroviral Vector ◽  
Human Chromosomes ◽  
G418 Resistance ◽  
Microcell Hybrid ◽  
Dominant Selectable Marker ◽  
Hybrid Clones ◽  
Microcell Fusion

We sought an efficient means to introduce specific human chromosomes into stable interspecific hybrid cells for applications in gene mapping and studies of gene regulation. A defective amphotropic retrovirus was used to insert the gene conferring G418 resistance (neo), a dominant selectable marker, into the chromosomes of diploid human fibroblasts, and the marked chromosomes were transferred to mouse recipient cells by microcell fusion. We recovered five microcell hybrid clones containing one or two intact human chromosomes which were identified by karyotype and marker analysis. Integration of the neo gene into a specific human chromosome in four hybrid clones was confirmed by segregation analysis or by in situ hybridization. We recovered four different human chromosomes into which the G418 resistance gene had integrated: human chromosomes 11, 14, 20, and 21. The high efficiency of retroviral vector transformation makes it possible to insert selectable markers into any mammalian chromosomes of interest.

Isolation of microcell hybrid clones containing retroviral vector insertions into specific human chromosomes

1987 ◽  
Vol 7 (8) ◽  
pp. 2814-2820
Author(s):  
T G Lugo ◽  
B Handelin ◽  
A M Killary ◽  
D E Housman ◽  
R E Fournier
Keyword(s):  
High Efficiency ◽  
Human Fibroblasts ◽  
Retroviral Vector ◽  
Human Chromosomes ◽  
G418 Resistance ◽  
Microcell Hybrid ◽  
Dominant Selectable Marker ◽  
Hybrid Clones ◽  
Microcell Fusion

We sought an efficient means to introduce specific human chromosomes into stable interspecific hybrid cells for applications in gene mapping and studies of gene regulation. A defective amphotropic retrovirus was used to insert the gene conferring G418 resistance (neo), a dominant selectable marker, into the chromosomes of diploid human fibroblasts, and the marked chromosomes were transferred to mouse recipient cells by microcell fusion. We recovered five microcell hybrid clones containing one or two intact human chromosomes which were identified by karyotype and marker analysis. Integration of the neo gene into a specific human chromosome in four hybrid clones was confirmed by segregation analysis or by in situ hybridization. We recovered four different human chromosomes into which the G418 resistance gene had integrated: human chromosomes 11, 14, 20, and 21. The high efficiency of retroviral vector transformation makes it possible to insert selectable markers into any mammalian chromosomes of interest.

scholarly journals Selective transfer of individual human chromosomes to recipient cells.

1985 ◽  
Vol 5 (1) ◽  
pp. 140-146 ◽  
Author(s):  
P J Saxon ◽  
E S Srivatsan ◽  
G V Leipzig ◽  
J H Sameshima ◽  
E J Stanbridge
Keyword(s):  
Hypoxanthine Phosphoribosyl Transferase ◽  
Human Chromosomes ◽  
Isoenzyme Analysis ◽  
Phosphoribosyl Transferase ◽  
Microcell Hybrid ◽  
Multiple Copies ◽  
G 11 ◽  
Hybrid Clones ◽  
Individual Human

Two hypoxanthine phosphoribosyltransferase-deficient human cell lines, D98/AH-2 and HT1080-6TG, were stably transfected with pSV2 gpt, a plasmid containing the selectable marker Escherichia coli xanthine-guanine phosphoribosyl transferase (Eco gpt). Hypoxanthine-aminopterin-thymidine-resistant transformants arose with a frequency of ca. 10(-6) and contained mostly single, but occasionally multiple, copies of the plasmid sequences. These transformants actively express the Eco gpt marker. Single chromosomes from two different HT1080 gpt transformants and one D98 gpt transformant, containing the integrated plasmid sequences, were transferred via microcell-mediated chromosome transfer to hypoxanthine phosphoribosyl transferase-deficient mouse A9 cells. The transferred human chromosomes were identified as 2, 4, and 22, by using a combination of G-11 staining, G-banding, isoenzyme analysis, and in situ hybridization. This system is being used to create a library of interspecies microcell hybrid clones, each clone containing a unique single human chromosome in a mouse background. The complete library will represent the entire human karyotype.

Selective transfer of individual human chromosomes to recipient cells

1985 ◽  
Vol 5 (1) ◽  
pp. 140-146
Author(s):  
P J Saxon ◽  
E S Srivatsan ◽  
G V Leipzig ◽  
J H Sameshima ◽  
E J Stanbridge
Keyword(s):  
Hypoxanthine Phosphoribosyl Transferase ◽  
Human Chromosomes ◽  
Isoenzyme Analysis ◽  
Phosphoribosyl Transferase ◽  
Microcell Hybrid ◽  
Multiple Copies ◽  
G 11 ◽  
Hybrid Clones ◽  
Individual Human

Two hypoxanthine phosphoribosyltransferase-deficient human cell lines, D98/AH-2 and HT1080-6TG, were stably transfected with pSV2 gpt, a plasmid containing the selectable marker Escherichia coli xanthine-guanine phosphoribosyl transferase (Eco gpt). Hypoxanthine-aminopterin-thymidine-resistant transformants arose with a frequency of ca. 10(-6) and contained mostly single, but occasionally multiple, copies of the plasmid sequences. These transformants actively express the Eco gpt marker. Single chromosomes from two different HT1080 gpt transformants and one D98 gpt transformant, containing the integrated plasmid sequences, were transferred via microcell-mediated chromosome transfer to hypoxanthine phosphoribosyl transferase-deficient mouse A9 cells. The transferred human chromosomes were identified as 2, 4, and 22, by using a combination of G-11 staining, G-banding, isoenzyme analysis, and in situ hybridization. This system is being used to create a library of interspecies microcell hybrid clones, each clone containing a unique single human chromosome in a mouse background. The complete library will represent the entire human karyotype.

The analysis of malignancy by cell fusion. VIII. Evidence for the intervention of an extra-chromosomal element

1977 ◽  
Vol 24 (1) ◽  
pp. 255-263
Author(s):  
J. Jonasson ◽  
H. Harris
Keyword(s):  
Human Chromosome ◽  
Gamma Radiation ◽  
Cell Fusion ◽  
Human Fibroblasts ◽  
Melanoma Cells ◽  
Human Chromosomes ◽  
Mouse Melanoma ◽  
Human Diploid Cells ◽  
Diploid Cells ◽  
Hybrid Clones

Diploid human fibroblasts and lymphocytes were fused with the cells of a malignant mouse melanoma and a range of hybrid clones selected for study. The ability of these clones to produce progressive tumours was assayed in nude mice. Although human chromosomes were preferentially eliminated in all the hybrid clones, the human diploid cells were as effective as mouse diploid cells in suppressing the malignancy of the mouse melanoma cells. The suppression produced by fibroblasts was again more profound than that produced by lymphocytes. Malignancy was also found to be suppressed in a hybrid clone in which a single X was the only human chromosome present; and this clone continued to give a very low take incidence even after the human X had been eliminated by back selection. Hybrids were made between the melanoma cells and diploid human fibroblasts that had been given 100 J kg-1 of gamma radiation before cell fusion. These hybrids contained no recognizable human chromosomes, but their ability to produce progressive tumours was greatly reduced compared to that of the melanoma parent cells. The take incidences given by the crosses between the melanoma cells and the irradiated human fibroblasts were, however, substantially higher than those given by the crosses between the melanoma cells and unirradiated fibroblasts. These findings suggest that the suppression of malignancy involves the activity of some extra-chromosomal element and that this element is radio-sensitive.

scholarly journals Influence of Casting Solvents on CO2/CH4 Separation Using Polysulfone Membranes

Membranes ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 286
Author(s):  
Roba M. Almuhtaseb ◽  
Ahmed Awadallah-F ◽  
Shaheen A. Al-Muhtaseb ◽  
Majeda Khraisheh
Keyword(s):  
High Efficiency ◽  
In Situ Synthesis ◽  
Separation Process ◽  
Polymeric Membranes ◽  
Gas Mixture ◽  
Manufacturing Cost ◽  
Binary Gas Mixture ◽  
Maximum Permeability ◽  
Polysulfone Membranes

Polysulfone membranes exhibit resistance to high temperature with low manufacturing cost and high efficiency in the separation process. The composition of gases is an important step that estimates the efficiency of separation in membranes. As membrane types are currently becoming in demand for CO2/CH4 segregation, polysulfone will be an advantageous alternative to have in further studies. Therefore, research is undertaken in this study to evaluate two solvents: chloroform (CF) and tetrahydrofuran (THF). These solvents are tested for casting polymeric membranes from polysulfone (PSF) to separate every single component from a binary gas mixture of CO2/CH4. In addition, the effect of gas pressure was conducted from 1 to 10 bar on the behavior of the permeability and selectivity. The results refer to the fact that the maximum permeability of CO2 and CH4 for THF is 62.32 and 2.06 barrer at 1 and 2 bars, respectively. Further, the maximum permeability of CF is 57.59 and 2.12 barrer at 1 and 2 bars, respectively. The outcome selectivity values are 48 and 36 for THF and CF at 1 bar, accordingly. Furthermore, the study declares that with the increase in pressure, the permeability and selectivity values drop for CF and THF. The performance for polysulfone (PSF) membrane that is manufactured with THF is superior to that of CF relative to the Robeson upper bound. Therefore, through the results, it can be deduced that the solvent during in-situ synthesis has a significant influence on the gas separation of a binary mixture of CO2/CH4.

Study on Mechanical Automation with Design of Rapid Milk Detector Based on Freezing Point

2013 ◽  
Vol 703 ◽  
pp. 282-286
Author(s):  
Ren Cai Zhang ◽  
Xiang Yu ◽  
Xing Ju Liu ◽  
Jin Hai Zhai ◽  
Zhen Wu Ning
Keyword(s):  
Integrated Circuit ◽  
Temperature Sensor ◽  
High Efficiency ◽  
Freezing Point ◽  
Electronic Control ◽  
Freezing Point Depression ◽  
Electronic Control System ◽  
Freezing Curve ◽  
Rack Mechanism

An efficient automated milk detector based on freezing point depression is designed. This detector shares characters of high efficiency and good stability with accuracy and automation. Its main parts include temperature sensor of IC (Integrated Circuit), pinion-rack mechanism and crank-rocker mechanism and electronic control system. Monitoring in-situ change of milk freezing curve and developing efficiency of sampling can be available by means of pinion-rack mechanism and IC temperature sensor mechatronics design. As a result, adulterating status of milk can be discriminated in a rapid and accurate and automated way. The detector may be employed to detect liquid foods other than milk as well.

Separation of phenols from oils using deep eutectic solvents and ionic liquids

2020 ◽  
Vol 92 (10) ◽  
pp. 1717-1731
Author(s):  
Yucui Hou ◽  
Zhi Feng ◽  
Jaime Ruben Sossa Cuellar ◽  
Weize Wu
Keyword(s):  
Ionic Liquids ◽  
Phenolic Compounds ◽  
High Efficiency ◽  
Deep Eutectic Solvents ◽  
Environmentally Benign ◽  
Organic Chemical ◽  
Environmental Friendliness ◽  
Oil Mixtures ◽  
Organic Chemical Industry

AbstractPhenolic compounds are important basic materials for the organic chemical industry, such as pesticides, medicines and preservatives. Phenolic compounds can be obtained from biomass, coal and petroleum via pyrolysis and liquefaction, but they are mixtures in oil. The traditional methods to separate phenols from oil using alkaline washing are not environmentally benign. To solve the problems, deep eutectic solvents (DESs) and ionic liquids (ILs) have been developed to separate phenols from oil, which shows high efficiency and environmental friendliness. In this article, we summarized the properties of DESs and ILs and the applications of DESs and ILs in the separation of phenols and oil. There are two ways in which DESs and ILs are used in these applications: (1) DESs formed in situ using different hydrogen bonding acceptors including quaternary ammonium salts, zwitterions, imidazoles and amides; (2) DESs and ILs used as extractants. The effect of water on the separation, mass transfer dynamics in the separation process, removal of neutral oil entrained in DESs, phase diagrams of phenol + oil + extractant during extraction, are also discussed. In the last, we analyze general trends for the separation and evaluate the problematic or challenging aspects in the separation of phenols from oil mixtures.

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