252 research outputs found
A new DNA-cloning vector for Haemophilus influenzae Rd
A new, high-efficiency, DNA-cloning vector pJ1-8 was derived in two steps from the chimeric plasmid pD7 consisting of RSF 0885(ampr) and Haemophilus influenzae chromosomal DNA. pJl-8 has only one EcoRI site and a molecular weight of only 2.5×106. No detectable ampr transformation was obtained with pJl-8 DNA. However, ampr transformation increases markedly if Haemophilus influenzae chromosomal DNA segments are spliced into it, providing a very facile assay for detecting inserts
Analysis of a genetically unstable region in Streptomyces lividans 66-TK64
Genetic and molecular analyses of an unstable region encompassing the gene loci cml arg and a 5.7 kb amplifiable unit of DNA were done. Spontaneous mutants from Cm1R→CmlS and the revertants from CmlS →CmlR were analysed for mutations at arg locus and amplification of amplifiable unit of DNA. Twenty-one revertants were analysed. Two of these had large-scale amplification and one of these was also Arg-. Nine of the revertants which were Arg+ had low-level or intermediate-level amplification of the 5.7 kb DNA sequence but no deletions of the flanking sequences were detected. Five of the CmIR' revertants, which were also Arg+, had lost one of the two copies from the doublet of amplifiable unit of DNA. The remaining five revertants did not show any other change. The amplifiable unit of DNA, therefore, not only undergoes amplification but can also suffer specific deletion of one copy. Thus, this region as a whole is characterized by instability and the events appear to take place at more than one locus concomitantly with a high frequency
Genetic transformation in bacteria
Certain species of bacteria can become competent to take up high molecular weight DNA from the surrounding medium. DNA homologous to resident chromosomal DNA is transported, processed and recombined with the resident DNA. There are some variations in steps leading to transformation between Gram-positive bacteria likebiplococcus pneumoniae and Gram-negative bacteria represented byHaemophilus influenzae but the integration is by single-strand displacement in both cases. Plasmid (RSF0885) transformation is low inHaemophilus influenzae but this is increased significantly if (homologous) chromosomal DNA is spliced to plasmid DNA. In Haemophilus influenzae, rec1 function is required for peak transformation with chimeric plasmids. Chimeric plasmid fixed presumably extrachromosomally undergoes frequent recombination between homologous segments contained in resident chromosome and the plasmid
An estimate of the physical distance between two linked markers in Haemophilus influenzae
Using DNA clones, the physical distance between the linked genesnov and str in Haemophilus influenzae was estimated. Although none of the cloned inserts contained both the markers, pJ1-8StrR 13 (insert of 18·7 kb) included str gene at one end and part of nov gene at the other end of the insert. By EcoRI restriction analysis and by Southern hybridization, the distance between the two EcoRI sites, cutting at which inactivates the two genes, was estimated to be 17·7 kb. A single continuous EcoRI fragment (containing 4EcoRI sites within it) carrying both the genes intact would need to be 20·4 kb in size. These estimates were confirmed independently using different clones of novr and strr alleles as probes for hybridization with BamHI-digested chromosomal DNA
Down-regulation of the global regulator SATB1 by statins in COLO205 colon cancer cells
Special AT-rich sequence binding protein 1 (SATB1) regulates the expression of more than 1,000 genes in tumor cells. SATB1 expression has been implicated in metastasis, and its silencing results in reduced cancer progression and the reversion of metastatic cells to normal appearance. Therefore, any compound causing down-regulation of SATB1 expression or activity may be exploited for its therapeutic potential in terms of cancer regression. Earlier studies showed that the 3-hydroxy-3-methylglutaryl coenzymeA (HMG-CoA) reductase inhibitors (statin drugs), which are widely used to treat hypercholesterolemia, possess other pleotropic activities. These are now increasingly gaining attention for their cancer prevention abilities. However, the downstream interplay of the molecular mechanisms of such anti-cancer activities is unclear. Here, we show that SATB1 is down-regulated by statins in a time- and dose-dependent manner in COLO205 cells. This effect was statin-specific as the down-regulation of SATB1 was brought about by hydrophobic statins, such as simvastatin and fluvastatin, but not by hydrophilic pravastatin. Notably, treatment with mevalonate, an intermediate in the cholesterol and isoprenoid biosynthetic pathways, led to the inhibition of SATB1 down-regulation and cytotoxicity mediated by statins. Treatment with the proteasome inhibitors lactacystine and MG-132 inhibited the statin-mediated down-regulation of SATB1, suggesting that regulation occurs at the post-translational level. Thus, our results demonstrate a novel molecular mechanism for the anti-cancer activity of statin drugs in colon cancer cells, without invoking significant cytotoxicity
Coulomb and nuclear breakup of a halo nucleus 11Be
Breakup reactions of the one-neutron halo nucleus 11Be on Pb and C targets at
about 70 MeV/u have been investigated by measuring the momentum vectors of the
incident 11Be, outgoing 10Be, and neutron in coincidence. The relative energy
spectra as well as the angular distributions of the 10Be+n center of mass have
been extracted for both targets. For the breakup on Pb target, the selection of
forward scattering angles is found to be effective to extract almost purely the
first-order E1 Coulomb breakup component, and to exclude the nuclear
contribution and higher-order Coulomb breakup components. This angle-selected
energy spectrum is thus used to deduce the spectroscopic factor for the
10Be(0+) 2s_1/2 configuration in 11Be which is found to be 0.72+-0.04 with
B(E1) up to Ex=4 MeV of 1.05+-0.06 e2fm2. The energy weighted E1 strength up to
Ex=4 MeV explains 70+-10% of the cluster sum rule, consistent with the obtained
spectroscopic factor. The non-energy weighted sum rule is used to extract the
root mean square distance of the halo neutron to be 5.77(16) fm, consistent
with previously known values. In the breakup with C target, we have observed
the excitations to the known unbound states in 11Be at Ex=1.78 MeV and 3.41
MeV. Angular distributions for these states show the diffraction pattern
characteristic of L=2 transitions, resulting in J^pi =(3/2,5/2)+ assignment for
these states. We finally find that even for the C target the E1 Coulomb direct
breakup mechanism becomes dominant at very forward angles.Comment: 14 pages, 7 figures, accepted for publication on Physical Review
Long-term carcinogenicity of pan masala in Swiss mice
Carcinogenicity of pan masala, a dry powdered chewing mixture of areca nut, catechu, lime, spices and flavoring agents was evaluated by means of the long-term animal bio-assay 6- to 7-week old male and female S/RVCri mice were divided randomly into intermediate and lifetime exposure groups and fed normal diet without pan masala - (zero dose) or diet containing 2.5% and 5% pan masala. Animals in the intermediate-exposure group (n = 10/gender/dose group) were killed after 6, 12 or 18 months of treatment, while those in the lifetime-exposure group (n = 54/gender/dose group) were killed when moribund or at the termination of the experiment at 24 months. Several tissues were processed for histopathological examination. The body weight and survival rate of mice fed pan masala were lower than that of the controls. Histopathological observations of tissues from control animals did not reveal any neoplastic alterations. However, lifetime feeding of pan masala induced adenoma of the liver, stomach, prostate and sebaceous glands, also forestomach papilloma, liver hamartoma, hepatoma and hemangioma, carcinoma of the forestomach, adenocarcinoma of the lung and liver, and testicular lymphoma. Neoplastic lesions appeared mainly in the liver (n = 13), stomach (n = 3) and lung (n = 8). Lung adenocarcinoma, the most frequent malignant tumor type, was observed in 2/120 mice in the intermediate-exposure group and in 8/216 animals in the lifetime-exposure group. Statistical analysis of tumor-induction data revealed a significant dose-related increase in lung adenocarcinomas but not in liver and stomach neoplasms indicating that lung is the major target tissue for the carcinogenic action of pan masala
Projectile fragmentation reactions and production of nuclei near the neutron drip-line
The reaction mechanism of projectile fragmentation at intermediate energies
has been investigated observing the target dependence of the production cross
sections of very neutron-rich nuclei. Measurement of longitudinal momentum
distributions of projectile-like fragments within a wide range of fragment mass
and its charge was performed using a hundred-MeV/n Ar beam incident on
Be and Ta targets. By measurement of fragment momentum distribution, a
parabolic mass dependence of momentum peak shift was observed in the results of
both targets, and a phenomenon of light-fragment acceleration was found only in
the Be-target data. The analysis of production cross sections revealed an
obvious enhancement of the target dependence except target size effect when the
neutron excess is increased. This result implies the breakdown of factorization
(BOF) of production cross sections for very neutron-rich nuclei near the drip
line.Comment: 16 pages, 18 figures, submitted to Phys. Rev.
Monohybrid and dihybrid segregations in the progenies of tobacco transformed for kanamycin resistance with a Ti-vector system
A chimeric DNA construction having nopaline synthase promoter, coding sequences of neomycin phosphotransferase gene conferring resistance to antibiotic kanamycin and OCS (octopine synthase) polyadenylation sequences bracketed by T-DNA ends was transferred to tobacco. Leaf discs were infected with A. tumefaciens containing disarmed, cointegrate plasmid pGV3850:: 1103 and allowed to form a callus in the presence of kanamycin. Shoots regenerated from infected leaf discs either through the callus or arising directly were further selected for their ability to root in kanamycin-containing media. Among the nine transgenic plants that were progeny tested, the transferred bacterial gene segregated as monohybrid ratio (3 KanR: 1 KanS) in seven. Segregation data of two plant progenies indicated the presence of two independent loci of KanR DNA insertion (15 KanR: 1 KanS). Back-cross segregation data were consistent with the monohybrid or independent assortment of duplicate factors. Thus in the two cases, a minimum independent integration of two copies of T-DNA each with a KanR marker is inferred
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