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11.
Gene transfer is a major factor in bacterial evolution 总被引:14,自引:3,他引:14
Lateral gene transfer in four strains of Salmonella enterica has been
assessed using genomic subtraction. Strain LT2 (subspecies I serovar
Typhimurium) chromosomal DNA was used as target and subtracted by three
subspecies I strains of serovars Typhimurium (S21), Muenchen (S71), Typhi
(M229), and a subspecies V strain (M321). Data from probing random cosmids
of LT2 DNA with preparations of the residual LT2 DNA after subtraction were
used to estimate the amounts of LT2 DNA not able to hybridize to strains
S21, S71, M229, and M321 to be in the range of 84-106, 191-355, 305-629,
and 778-1,286 kb, respectively. Several lines of evidence indicate that
most of this DNA is from genes not present in strain M321 and not from
genes that have diverged in sequence. The amounts correlate with the
divergence of the four strains as revealed by multilocus enzyme
electrophoresis and sequence variation of housekeeping genes. Sequence of
39 of the fragments from the M321 subtracted residual LT2 DNA revealed only
six inserts of known gene function with evidence of both gain and loss of
genes during the development of S. enterica clones. Sixteen of the 39
segments have 45% or lower G+C content, below the species average, but over
half are within the normal range for the species. We conclude that even
within a species, clones may differ by up to 20% of chromosomal DNA,
indicating a major role for lateral transfer, and that on the basis of G+C
content, a significant proportion of the DNA is from distantly related
species.
相似文献
12.
为了研究禽流感H5N1病毒在各个器官的增殖和病理变化,在生物安全实验室,我们将禽流感H5N1病毒通过尾静脉接种BALB/C小鼠。结果小鼠在不经过适应的情况下,直接感染发病,甚至死亡。在观察的7天内,感染小鼠临床症状主要表现呼吸急促,体温、体重下降。尸检表现肺出血,心外膜坏死以及肝脏的坏死。组织病理检查表现心、肝、肺等多器官的病变。肺的病变伴有纤维化的弥漫性肺泡损伤;心肌外膜大量淋巴细胞浸润、坏死;肝细胞大量坏死,淋巴细胞浸润。心、肝的坏死病变在H5N1禽流感病毒相关的研究中未见报道。经过对各个组织器官的病毒载量的检测,未发现病毒在各个病变组织中的复制。免疫组化的检测,各个组织中也未检出阳性的细胞反应。因此,我们认为H5N1禽流感病毒感染小鼠引起多个器官组织的损伤,甚至死亡,不是病毒在器官的复制,而可能是病毒感染小鼠,产生炎症细胞因子的高度表达,损伤多个器官组织所致。 相似文献
13.
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15.
N-Linked glycosylation of a baculovirus-expressed recombinant glycoprotein in insect larvae and tissue culture cells 总被引:5,自引:3,他引:2
The potential of insect cell cultures and larvae infected with recombinant
baculoviruses to produce authentic recombinant glycoproteins cloned from
mammalian sources was investigated. A comparison was made of the N-linked
glycans attached to secreted alkaline phosphatase (SEAP) produced in four
species of insect larvae and their derived cell lines plus one additional
insect cell line and larvae of one additional species. These data survey
N-linked oligosaccharides produced in four families and six genera of the
order Lepidoptera. Recombinant SEAP expressed by recombinant isolates of
Autographa californica and Bombyx mori nucleopolyhedroviruses was purified
from cell culture medium, larval hemolymph or larval homogenates by
phosphate affinity chromatography. The N-linked oligosaccharides were
released with PNGase-F, labeled with 8- aminonaphthalene-1-3-6-trisulfonic
acid, fractionated by polyacrylamide gel electrophoresis, and analyzed by
fluorescence imaging. The oligosaccharide structures were confirmed with
exoglycosidase digestions. Recombinant SEAP produced in cell lines of
Lymantria dispar (IPLB-LdEIta), Heliothis virescens (IPLB-HvT1), and Bombyx
mori (BmN) and larvae of Spodoptera frugiperda, Trichoplusia ni ,
H.virescens , B.mori , and Danaus plexippus contained oligosaccharides that
were structurally identical to the 10 oligosaccharides attached to SEAP
produced in T.ni cell lines. The oligosaccharide structures were all
mannose-terminated. Structures containing two or three mannose residues,
with and without core fucosylation, constituted more than 75% of the
oligosaccharides from the cell culture and larval samples.
相似文献
16.
Plasma from hibernating woodchucks was desalted utilizing a hollow fiber device having a M. W. cut-off of 5,000. This preparation was fractionated by isoelectric focusing (IEF) in a pH gradient extending from 3.5 to 10.0 resulting in protein components having isoelectric points (pIs) of 4.5, 5.2, 5.5, 6.3, and 7.0. Fraction I (comprised of proteins having pIs of 4.5 and 5.2) induced hibernation within 2 to 6 days in 8 out of 10 summer-active ground squirrels. Fraction II (pI 5.5) and Fraction III (pI 6.3 and 7.0) failed to induce any summer hibernation in 10 animal test groups at identical sample concentrations. Polyacrylamide gel electrophoresis of Fraction I indicated that albumin was a major constituent of this still heterogeneous preparation. Thus, in order to more clearly define the plasma locus of this hibernation inducing trigger(s) (HIT) molecule, whole plasma and/or Fraction I was fractionated by 3 distinct resolving techniques. These included sub-fractionation of Fraction I by isoelectric focusing utilizing a narrower pH gradient extending from 3.5 to 6.0, isotachophoresis of whole plasma and affinity chromatography of Fraction I and whole plasma. A total of 40 summer-active ground squirrels were injected and assayed for HIT activity with fractionated preparations derived by the three previously cited separation techniques. A total of 18 of these summer-active ground squirrels hibernated. However, a much more impressive figure is that 16 out of 21 animals hibernated when injected with resolved hibernating plasma fractions in which albumin was the predominant plasma protein. A total of 8 control animals were injected with vehicle and none of these hibernated. 相似文献
17.
In situ hybridization at the electron microscope level: hybrid detection by autoradiography and colloidal gold 总被引:14,自引:4,他引:10 下载免费PDF全文
In situ hybridization has become a standard method for localizing DNA or RNA sequences in cytological preparations. We developed two methods to extend this technique to the transmission electron microscope level using mouse satellite DNA hybridization to whole mount metaphase chromosomes as the test system. The first method devised is a direct extension of standard light microscope level using mouse satellite DNA hybridization to whole mount metaphase chromosomes as the test system. The first method devised is a direct extension of standard light microscope in situ hybridization. Radioactively labeled complementary RNA (cRNA) is hybridized to metaphase chromosomes deposited on electron microscope grids and fixed in 70 percent ethanol vapor; hybridixation site are detected by autoradiography. Specific and intense labeling of chromosomal centromeric regions is observed even after relatively short exposure times. Inerphase nuclei present in some of the metaphase chromosome preparations also show defined paatterms of satellite DNA labeling which suggests that satellite-containing regions are associate with each other during interphase. The sensitivity of this method is estimated to at least as good as that at the light microscope level while the resolution is improved at least threefold. The second method, which circumvents the use of autoradiogrphic detection, uses biotin-labeled polynucleotide probes. After hybridization of these probes, either DNA or RNA, to fixed chromosomes on grids, hybrids are detected via reaction is improved at least threefold. The second method, which circumvents the use of autoradiographic detection, uses biotin-labeled polynucleotide probes. After hybridization of these probes, either DNA or RNA, to fixed chromosomes on grids, hybrids are detected via reaction with an antibody against biotin and secondary antibody adsorbed to the surface of over centromeric heterochromatin and along the associated peripheral fibers. Labeling is on average ten times that of background binding. This method is rapid and possesses the potential to allow precise ultrastructual localization of DNA sequences in chromosomes and chromatin. 相似文献
18.
Leonid N. Maslov Yury B. Lishmanov Peter R. Oeltgen Eva I. Barzakh Andrey V. Krylatov Meera Govindaswami Stephen A. Brown 《Life sciences》2009,84(19-20):657-663
AimsThis study aims to investigate the role of peripheral δ2 opioid receptors in cardiac tolerance to ischemia/reperfusion injury and to examine the contribution of PKC, TK, KATP channels and the autonomic nervous system in δ2 cardioprotection.Main methodsDeltorphin II and various inhibitors were administered in vivo prior to coronary artery occlusion and reperfusion in a rat model. The animals were monitored for the development of arrhythmias, infarct development and the effects of selected inhibitors.Key findingsPretreatment with peripheral and δ2 specific opioid receptor (OR) antagonists completely abolished the cardioprotective effects of deltorphin II. In contrast, the selective δ1 OR antagonist 7-benzylidenenaltrexone (BNTX) had no effect. The protein kinase C (PKC) inhibitor chelerythrine and the NO-synthase inhibitor L-NAME (N-nitro-l-arginine methyl ester) also reversed both deltorphin II effects. The nonselective ATP-sensitive K+ (KATP) channel inhibitor glibenclamide and the selective mitochondrial KATP channel inhibitor 5-hydroxydecanoic acid only abolished the infarct-sparing effect of deltorphin II. Inhibition of tyrosine kinase (TK) with genistein, the ganglion blocker hexamethonium and the depletion of endogenous catecholamine storage with guanethidine reversed the antiarrhythmic action of deltorphin II but did not change its infarct-sparing action.SignificanceThe cardioprotective mechanism of deltorphin II is mediated via stimulation of peripheral δ2 opioid receptors. PKC and NOS are involved in both its infarct-sparing and antiarrhythmic effects. Infarct-sparing is dependent upon mitochondrial KATP channel activation while the antiarrhythmic effect is dependent upon TK activation. Endogenous catecholamine depletion reduced antiarrhythmic effects but did not alter the infarct-sparing effect of deltorphin II. 相似文献
19.
Yu. B. Lishmanov L. N. Maslov T. V. Lasukova A. A. Platonov P. Oeltgen 《Biology Bulletin》2005,32(1):45-51
Activation of cardiac -opioid receptors (ORs) by their selective agonist DPDPE (154 nM) increased the resistance of perfused rat heart to ischemia/reperfusion injury. Decreased release of creatine phosphokinase to the perfusate and decreased incidence of arrhythmias were observed during reoxygenation. At the same time, opioidergic decrease in left ventricular developed pressure took place both during the preischemic period and after restoring the coronary circulation. All these effects could be prevented by blocking -ORs by naltrindole or inhibition of Ca2+-ATPase of sarcoplasmic reticulum by cyclopiazonic acid. -OR agonist DPDPE had no effect on cAMP levels in myocardial tissue during the whole experiment. The obtained data suggest that the antiarrhythmic and cytoprotective effects observed after -OR stimulation can be realized through the changes in Ca2+ transport at the level of the sarcoplasmic reticulumTranslated from Izvestiya Akademii Nauk, Seriya Biologicheskaya, No. 1, 2005, pp. 55–62.Original Russian Text Copyright © 2005 by Lishmanov, Maslov, Lasukova, Platonov, Oeltgen. 相似文献
20.
P. R. Oeltgen L. C. Bergmann W. A. Spurrier S. B. Jones 《Preparative biochemistry & biotechnology》2013,43(2-3):171-188
Plasma from hibernating woodchucks was desalted utilizing a hollow fiber device having a M. W. cut-off of 5, 000. This preparation was fractionated by isoelectric focusing (IEF) in a pH gradient extending from 3. 5 to 10. 0 resulting in protein components having isoelectric points (pis) of 4. 5, 5. 2, 5. 5, 6. 3, and 7. O. Fraction I (comprised of proteins having pis of 4. 5 and 5. 2) induced hibernation within 2 to 6 days in 8 out of 10 summer-active ground squirrels. Fraction II (pI 5. 5) and Fraction III (pi 6. 3 and 7. 0) failed to induce any summer hibernation in 10 animal test groups at identical sample concentrations. Polyacrylamide gel electrophoresis of Fraction I indicated that albumin was a major constituent of this still heterogeneous preparation. Thus, in order to more clearly define the plasma locus of this hibernation inducing trigger(s) (HIT) molecule, whole plasma and/or Fraction I was fractionated by 3 distinct resolving techniques. These included sub-fractionation of Fraction I by isoelectric focusing utilizing a narrower pH gradient extending from 3. 5 to 6. 0, isotachophoresis of whole plasma and affinity chromatography of Fraction I and whole plasma. A total of 40 summer-active ground squirrels were injected and assayed for HIT activity with fractionated preparations derived by the three previously cited separation techniques. A total of 18 of these summer-active ground squirrels hibernated. However, a much more impressive figure is that 16 out of 21 animals hibernated when Injected with resolved hibernating plasma fractions in which albumin was the predominant plasma protein. A total of 8 control animals were injected with vehicle and none of these hibernated. 相似文献