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C J Collins

Publications and source records attributed to C J Collins.

26 records · Page 2Linked to original sources

Effects of chronic inhalation of dimethyl ether in the rat.

Dimethyl ether (DME) is a colourless gas with an ethereal odour. It is used mainly as a liquid refrigerant and an aerosol dispersant. Little or no data exist on the toxicology of this material. In the present study, groups of male and female rats were exposed to 0.02%, 0.2% and 2% v/v of DME in air, 6 h/day, 5 days/week for 30 weeks. At the end of the study the high-dose level male rats showed a significant reduction on liver weight compared with the control group; this was accompanied by raising SGPT levels. In high-dose female rats there was no significant effect on liver weight but the SGPT levels were raised above control values. No histological abnormalities were observed in the liver or in any other organ.

Aerosols↗

Integration of avian sarcoma virus DNA sequences in transformed mammalian cells.

DNA from six avian sarcoma virus (ASV)-transformed mammalian cell lines was digested with the restriction endonucleases EcoRI, Xho I, or Sal I, fractionated by agarose gel electrophoresis, transferred to nitrocellulose filter strips, and hybridized with specific ASV [32P]cDNA probes. DNA from all of the ASV-transformed cell lines yielded three common virus-specific DNA fragments (2.4, 1.8, and 1.3 X 10(6) daltons) upon cleavage with EcoRI. Xho I appeared to cleave at least once within the integrated provirus and yielded a common fragment of 3.3 X 10(6) daltons as well as a second virus-specific DNA fragment whose size varied from 4.0 to 5.0 X 10(6) daltons in the different transformed cell lines. Sal I did not cleave within the provirus and yielded a single major virus-specific fragment of about 11 X 10(6) daltons in all transformed lines examined. Using specific cDNA probes, we show that the 1.8 X 10(6)-dalton EcoRI fragment contains sequences homologous to the 3' end of the viral RNA as well as to the src region of the viral genome. These studies clearly demonstrate that the same region on the ASV genome is utilized for provirus integration in different ASV-transformed cell lines.

Avian Sarcoma Viruses↗

Joining of simian virus 40 DNA molecules at endonuclease R Eco Ri sites by polynucleotide ligase and analysis of the products by agarose gel electrophoresis.

DNA molecules cut with endonuclease R Eco Ri can be joined at Eco Ri cleavage sites by incubation with polynucleotide ligase. In order to define the optimum conditions for this reaction, linear Simian Virus 40 DNA molecules (SV40(Lri)) produced by endonuclease R Eco Ri cleavage of SV40 form i DNA were joined using polynucleotide ligases specified by bacteriophage T4 and Escherichia coli. We have determined that the concentration of the substrate DNA molecules is the most important factor determining the distribution of covalently joined product molecules into a variety of circular and linear monomeric and oligomeric species.

DNA Restriction Enzymes↗

Biochemical construction of specific chimeric plasmids from ColE1 DNA and unfractionated Escherichia coli DNA.

A series of chimeric plasmids was constructed using colicinigenic factor E1 (ColE1) DNA as the replicon and DNA fragments carrying the galactose or tryptophan operons from E. coli. Restriction endonuclease EcoRI digests of ColE1 DNA and various DNAs containing the trp or gal operons were joined by T4 polynucleotide ligase [polynucleotide synthetase (ATP), poly(deoxyribonucleotide):poly(deoxyribonucleotide) ligase (AMP-forming), EC 6.5.1.1]. Chimeric plasmids carrying the desired genes were selected after transformation of Trp- or Gal- cells with ligated DNA. By using this method, we constructed ColE1-gal and ColE1-trp chimeric plasmids in which the source of the bacterial gal and trp operons was an unfractionated EcoRI digest of total E. coli DNA. The frequency of recovery of such chimeric plasmids is 10 to 20 colonies per mug of ligated DNA used in the transformation step. The method utilized in this report for constructing specific chimeric plasmids from total E. coli DNA is very simple. It requires only endonuclease R-EcoRI and T4 polynucleotide ligase, both of which are commercially available. The yield of transformants suggests that this method will be useful for cloning and amplifying a wide variety of functionally defined genes from E. coli and other prokaryotic organisms.

Colicins↗

Events occurring near the time of synapsis during transformation in Diplococcus pneumoniae.

A marker-specific and strongly temperature-dependent reaction was observed to occur at a time during transformation in Diplococcus pneumoniae after the donor deoxyribonucleic acid (DNA) had acquired single-strand properties and immediately preceding the integration of these strands into the recipient chromosome. Operationally, it was observed as the prevention of an intracellular inactivation process, also described in this paper, which is specific for low molecular weight or for damaged DNA, and which occurs if the recipient cells are held at suboptimal temperatures after the DNA has entered. Brief exposure of the cells to a higher temperature stabilized the DNA against this inactivation, in a two step process. It is the first step which has a strong temperature dependence (DeltaHdouble dagger = 70 kcal/mole, DeltaSdouble dagger = 160 entropy units), is marker specific, and which appears to be reversible. The second step is much less temperature-dependent and overlaps in time the start of integration. The enthalpy and entropy of activation are both consistent with those needed to open a loop of six to eight base pairs in a DNA duplex. It is suggested that these observations may reflect, and provide an assay for, the kinetics of synapsis, which on this model is limited in rate by the appearance of unpaired regions on the recipient duplex.

Chromosomes, Bacterial↗

Fate of infecting simian virus 40-deoxyribonucleic acid in nonpermissive cells: integration into host deoxyribonucleic acid.

Nonpermissive 3T3 cells were infected with purified superhelical simian virus 40 (SV40) deoxyribonucleic acid I (DNA I). One hour after infection, approximately 60% of the intracellular SV40 DNA was converted to relaxed forms. One day after infection, all intracellular SV40 DNA was present as slow-sedimenting material, and no SV40 DNA I was detectable. At 2 days after infection there appeared viral DNA sequences cosedimenting with cellular DNA during alkaline velocity centrifugation. Furthermore, by both alkaline equilibrium gradient centrifugation and by DNA-ribonucleic acid hybridization analysis, covalent linkage of viral DNA sequences to cellular DNA was demonstrated. Integration of SV40 DNA into cellular DNA did not appear to require DNA synthesis, although DNA synthesis followed by mitotic division of the cells enhanced the amount of viral DNA integrated. Based on data obtained by two different methods, it was calculated that 1,100 to 1,200 SV40 DNA equivalents must be integrated per cell by 48 hr after infection.

Adsorption↗