Gene/Protein Disease Symptom Drug Enzyme Compound
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After transfer from a mutagenized host, twenty one Co1E2 plasmid mutants were isolated after screening 10,000 clones for abnormal colicin production. Analysis by SDS polyacrylamide slab gel electrophoresis of proteins synthesized after mitomycin C-induction of mutant cultures, indicates that all but two of the mutations are in the structural gene from colicin E2. Of these, nine produce fragments of colicin in both whole cells and minicells and some are suppressed by nonsense suppressors. Studies with a nonsense mutant producing only a small colicin E2 fragment (Co1E2-421) suggest that colicin E2 in not involved in plasmid DNA replication, in the control of its own synthesis, or required for cell death when cells become committed to colicin production. The two plasmid mutants outside the colicin gene segregate plasmid-free cells at 33 degrees, 37 degrees and 43 degrees. One segregates fairly rapidly (about 4% per generation) though the colicin-producing cells make normal amounts of colicin, whilst the other segregates more slowly and the colicin-producing cells make much reduced amounts of colicin.
Mol Gen Genet 1976 Aug 02
PMID:Isolation and characterization of Co1E2 plasmid mutants unable to kill colicin-sensitive cells. 79 89

Aspergillus foetidus virus S (AfV-S) and A. foetidus virus F(AfV-F) have been shown to be serologically unrelated. Amino acid compositions of the two virus capsids are compared and their capsid polypeptides have been examined by SDS-polyacrylamide gel electrophoresis. AfV-F contained one major (phi3) and two minor (phi1 and phi2) polypeptides with mol. wt. 87000, 125000 and 100000, while AfV-S contained one major (sigma1) and one minor (sigma2) polypeptide with mol. wt. 83000 and 78000 respectively. Evidence is presented that sigma2 may be derived from sigma1 polypeptide by proteolytic degradation in vitro. The mol. wt. of AfV-F4 and AfV-S1a particles were found from sedimentation and diffusion coefficients to be 13 I times 10-6 and 12 4 times 10-6 respectively. AfV-F capsid was estimated to contain 120 molecules of polypeptide phi3 and one molecule each of polypeptides phi1 and phi2, while AfV-S capsid was estimated to contain 120 molecules of polypeptide phi1. It has been shown that SIa and S-2a particles each contain a molecule of double-stranded RNA with mol. wt. 2 24 times 10-6 (RNA-224) and 2 76 times 10-6 (RNA-276) respectively, whereas S-Ib and S-2b particles each contain a molecule of RNA-224 and RNA-276 respectively, together with an additional molecule of double-stranded RNA of mol. wt. 0 I times 10-6. Evidence is presented that S4 particles contain two molecules of RNA-224. S3 particles gave only RNA-224 on extraction, but contain the equivalent of 11/2 molecules of RNA-224; the nature of these particles and other possible virus replicative intermediates is discussed. Double-stranded RNA of mol. wt. I 24 times 10-6 was derived from a newly described particle class, Fo.
J Gen Virol 1975 May
PMID:Biophysical and biochemical properties of two viruses isolated from Aspergillus foetidus. 80 61

Bacteriophage MX-1 is a virulent DNA phage for Myxococcus. The host range includes strains of Myxococcus xanthus, M. fulvus and M. virescens. The phage has a sedimentation coefficient (S degrees 20,w) of 1145S and a density of 1-531 g/ml. By using SDS-polyacrylamide gel electrophoresis, 23 phage proteins with apparent mol. wt. between 10000 and 150000 were resolved. Gel filtration in the presence of non-ionic detergent partially resolved the proteins. The fraction excluded from Sephadex G-100, fraction 1, contains two glycoproteins. Fraction 1 was resolved into three fractions (1-1, 1-2 and 1-3) by chromatography on Sephadex G-200. The glycoproteins were present in fraction 1-2; all the proteins from this fraction were derived from the phage tail. Comparison of the amino-acid, hexosamine and neutral-sugar compositions of the two glycoproteins showed that they are distinct molecular species; the smaller molecule is not a subunit of the larger. The significance of these findings is discussed and compared with the proteins of the tails of T-even phage of Escherichia coli.
J Gen Virol 1976 Jan
PMID:Bacteriophage MX-1: properties of the phage and its structural proteins. 81 57

A total of 23 phage specific proteins (including four head and six tail proteins) could be identified after SDS polyacrylamide gel electrophoresis of extracts from phage SPP1 infected Bacillus subtilis cells. The total molecular weight of the proteins amounts to approximately 1.9 X 10(6) daltons, equivalent to the majority of the coding capacity of SPP1 DNA. It can thus be assumed that almost all SPP1 coded proteins have been identified. Protein assignments to phage cistrons were made by analysis of extracts from nonpermissive cells infected with sus-mutants. The SPP1 specified proteins can be subdivided into three groups on the basis of the time of their synthesis during the latent period. Host protein synthesis is not significantly affected by SPP1 infection. Normal expression of host genes appears to be essential for SPP1 growth.
Mol Gen Genet 1975 Dec 23
PMID:Gene expression of bacteriophage SPPI. I. Phage directed protein synthesis. 81 1

The biophysical and biochemical properties of Penicillium cyaneo-fulvum virus (Pc-fV) and Penicillium chrysogenum virus (PcV) have been compared. In sucrose density gradient sedimentation, purified virus preparations gave one major component, L, and three minor components E1, E2 and H with sedimentation coefficients of 145S, 80S, 102S and 172S respectively in each case. E1, E2 were shown to be empty particles. Pc-fV L particles contained only double-stranded RNA, which separated in polyacrylamide gel electrophoresis into four components with mol. wt. 2-21 X 10(6), 2-08 X 10(6), 1-98 X 10(6) and 1-93 X 10(6). PcV L particles gave three double-stranded RNA components, which were indistinguishable in polyacrylamide gel electrophoresis from the larger three RNA components of Pc-fV. In both viruses each RNA component was separately encapsidated. In both viruses H particles gave rise to the same double-stranded RNA components as their L particles, but contained, in addition, small amounts of single-stranded RNA. Pc-fV and PcV have been shown to consist of isometric particles of the same size and to be serologically related, and the amino acid compositions of their capsid polypeptides were found to be very similar. The capsid polypeptides of the two viruses were examined by SDS-polyacrylamide gel electrophoresis. In each case E2, L and H particles gave one major polypeptide gamma1, with mol. wt. 130000, whereas E1 particles contained one major polypeptide gamma2 with mol. wt. 115000. The mol. wt. of L particles, determined from sedimentation and diffusion coefficients, was 9-8 X 10(6) for both Pc-fV and PcV. The capsid of the L particles of each virus was estimated to contain 60 molecules of polypeptide gamma1.
J Gen Virol 1977 Jan
PMID:Comparison of the biophysical and biochemical properties of Penicillium cyaneo-fulvum virus and Penicillium chrysogenum virus. 83 75

The structural polypeptides of five bunyaviruses, snowshoe hare, Lumbo and La Crosse viruses (members of the California encephalitis subgroup of bunyaviruses), Bunyamwera and Main Drain viruses (members of the Bunyamwera subgroup of bunyaviruses), have been compared by polyacrylamide-SDS gel electrophoresis. Each virus was found to possess three major structural polypeptides, two glycoproteins (G1 and G2), and one nucleocapsid protein (N). Although the sizes of the G1 polypeptides (mol. wt. approx. 115 X 10(3)) and G2 polypeptides (mol. wt. approx. 38 X 10(3)) of the five viruses were found to be essentially similar, the sizes of the N polypeptides of the various viruses differed (mol. wt. range 19 to 24 X 10(3)). The RNA genomes of four bunyaviruses (snowshoe hare, La Crosse, Bunyamwera and Main Drain) have also been compared. Each virus has three RNA species of mol. wt. approx. 3 X 10(6), 1-9 X 10(6) and 0-4 X 10(6). Minor size differences were observed for the smallest RNA species of the four viruses (mol. wt. range 0-34 to 0-50 X 10(6)). For snowshoe hare virus the RNA segments hav a 5' sequence of pppAp...which suggests that the RNA is linear and not circular.
J Gen Virol 1977 Feb
PMID:The virus particle nucleic acids and proteins of four bunyaviruses. 83 99

Human embryonic kidney cells have been transformed by exposing cells to sheared fragments of adenovirus type 5 DNA. The transformed cells (designated 293 cells) exhibited many of the characteristics of transformation including the elaboration of a virus-specific tumour antigen. Analysis of the polypeptides synthesized in the 293 cells by labelling with 35S-methionine and SDS PAGE showed a variable pattern of synthesis, different in a number of respects from that seen in otheruman cells. On labelling the surface of cells by lactoperoxidase catalysed radio-iodination, the absence of a labelled polypeptide analogous to the 250 K (LETS) glycoprotein was noted. Hybridization of labelled cellular RNA with restriction fragments of adenovirus type 5 DNA indicated transcription of a portion of the adenovirus genome at the conventional left hand end.
J Gen Virol 1977 Jul
PMID:Characteristics of a human cell line transformed by DNA from human adenovirus type 5. 88 4

The matrix (M) protein of the H2N2 virus A/Ann Arbor/6/60 may be distinguished from M protein of several H3N2 viruses and A/New Jersey/76 (HSWINI) by SDS acrylamide gel electrophoresis using a discontinuous buffer system. The smallest RNA (RNA 8) of the A/Ann Arbor/6/60 virus may be distinguished from RNA 8 of several H3N2 viruses by acrylamide gel electrophoresis in 3% or 3-6% gels in the absence of urea, if electrophoresis is done at 30 to 36 degrees C or 20 degrees C respectively. Ten clones of conditionally-lethal temperature-sensitive (ts) mutants were studied, which derived their cold-adaption and ts genes from mutant A/Ann Arbor/6/60, and their haemagglutinin from the H3N2 virus A/Scotland/840/74. Each clone was found to derive its M protein from A/Ann Arbor/6/60 mutant, and its RNA 8 from A/Scotland/840/74. The only assignment of genes 7 and 8 consistent with these findings for the recombinants is that in each parent virus (and in the recombinants) gene 7 codes for M protein, and gene 8 for NS protein. Furthermore, it may be concluded from the results that the biologically important ts lesions in the A/Ann Arbor/6/60 mutant parent are not present in the NS gene. In addition to the recombinants of A/Ann Arbor/6/60 and A/Scotland/840/74, five independent ts/cold-adapted recombinants of A/Ann Arbor/6/60 mutant with H3N2 and HSWINI wild-type viruses were examined, and all were found to contain the M protein of the A/Ann Arbor/6/60 mutant parent. This is suggestive that M protein may be at least partially responsible for the cold-adaptation and/or ts properties of the A/Ann Arbor/6/60 mutant and the recombinants.
J Gen Virol 1977 Oct
PMID:Comparative studies of wild-type and 'cold-mutant' (temperature sensitive) influenza viruses: geneology of the matrix (M) and non-structural (NS) proteins in recombinant cold-adapted H3N2 viruses. 91 81

It has been determined that the bag cells of Aplysia californica produce two polypeptide species that comigrate on electrophoretic gels containing sodium dodecyl sulfate. By this separation procedure both species can be assigned a molecular weight of approximately 6,000. One of these molecules has an Rf of 0.65 on alkaline discontinuous electrophoresis gels, an isoelectric point at pH 4.8, a gel filtration molecular weight of approximately 12,000, and has no known biological function. The other does not enter alkaline disk gels, has an isoelectric point at approximately pH 9.3, shows a gel filtration molecular weight consistent with that determined by SDS gel electrophoresis, and is the egg-laying hormone.
J Gen Physiol 1976 Aug
PMID:Biochemical isolation and physiological identification of the egg-laying hormone in Aplysia californica. 95 70

When the polypeptides of hepatitis B surface antigen were examined by SDS-polyacrylamide gel electrophoresis under a variety of conditions, anomalous results were found to be due to (i) variable and at times incomplete dissociation of polypeptides after boiling with 1% SDS and reducing agent, (ii) reaggregation of solubilized material under certain electrophoretic conditions and during laboratory manipulations, and (iii) the variable presence of additional components in hepatitis B surface antigen prepared from certain individual donors. When these factors were taken into account, two major components were consistently identified by discontinuous buffer polyacrylamide gel electrophoresis, of apparent mol. wt. 60000 to 70000 and 12000 to 14000. However, in view of the demonstrated limitations of this technique in examining HBsAg polypeptides, alternative methods are necessary to confirm the true mol. wt. of the unique virus-specified amino acid sequence present.
J Gen Virol 1976 Nov
PMID:Examination of the polypeptides of hepatitis B surface antigen. 99 89


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