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101.
Quezada-Rivera JJ RE Soria-Guerra FS Pérez-Juárez L Martínez-González SE Valdés- Rodríguez NL Vasco-Méndez JF Morales-Domínguez 《Phyton》2019,88(1):25-35
The use of antimicrobial peptides (AMPs) synthesized
by bacteria (bacteriocins) is an alternative for combating multidrug
resistant bacterial strains and their production by recombinant route
is a viable option for their mass production. The bacteriocin E-760
isolated from the genus Enterococcus sp. has been shown to possess
inhibitory activity against Gram-negative and Gram-positive
bacteria. In this study, the expression of a chimeric protein coding
for E-760 in the nucleus of C. reinhardtii was evaluated, as well as,
its antibacterial activity. The synthetic gene E-760S was inserted
into the genome of C. reinhardtii using Agrobacterium tumefaciens.
A transgenic line was identified in TAP medium with hygromycin
and also by PCR. The increment in the culture medium temperature
of the transgenic strain at 35 °C for 10 minutes, increased the
production level of the recombinant protein from 0.14 (Noninduced
culture, NIC) to 0.36% (Induced culture, IC) of total soluble
proteins (TSP); this was quantified by an ELISA assay. Recombinant
E-760 possesses activity against Staphylococcus aureus in 0.34 U
log, Streptococcus agalactiae in 0.48 U log, Enterococcus faecium in
0.36 U log, Pseudomonas aeruginosa in 2 U log and for Klebsiella
pneumoniae, the activity was 0.07 U log. These results demonstrate
that the nucleus transformation of C. reinhardtii can function as
a stable expression platform for the production of the synthetic
gene E-760 and it can potentially be used as an antibacterial agent. 相似文献
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The events leading to the completion of cytokinesis after the formation of the midbody and intercellular bridge in D-98S cells were studied with light and electron microscopy. Pairs of daughter cells corresponding to different stages of cytokineses, as determined previously form time lapse films, were selected from embedded monolayers for serial sectioning. Separation of daughter cells is preceded by the reduction in diameter of the intercellular bridge from 1-1.5 μm to approx. 0.2 μm. Two processes contribute to this reduction: (a) The intercellular bridge becomes gradually thinner after telophase; a progressive breakdown of midbody structures accompanies this change; and (b) the more significant contribution to reduction in bridge diameter occurs through the localized constriction of a segment of the intercellular bridge.. The microtubules within the constricted portion of the bridge are forced closer together, and some microtubules disappear as this narrowing progresses. The plasma membrane over the narrowed segments is thrown into a series of wavelike ripples. Separation of daughter cells is achieved through movements of the cells which stretch and break the diameter-reduced bridge. The midbody is discarded after separation and begins to deteriorate. Occasional pairs of daughter cells were found in which incomplete karyokineses resulted in their nuclei being connected by a strand of nuclear material traversing the bridge and midbody. Such cells do not complete cytokinesis but merge together several hours after telophase. This merging of daughter cells coincides with the nearly complete breakdown of the midbody. 相似文献
106.
Secretion and degradation of parathormone as a function of intracellular maturation of hormone pools
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The biosynthesis, processing, and secretion of parthormone and the effect of calcium on these processes were measured in dispersed porcine parthyroid cells incubated with [(35)S]methionine. Proparathormone was detected at 10 min, the earliest time measured, and was rapidly and apparently quantitatively converted to parathormone. The half-life of the prohomormone pool was 15 min. Secretion of parathormone was detected by 20 min. In pulse-chase experiments there was a period between 20 and 40 min during which the wave of newly-synthesized parathormone was secreted. After 40 min during little additional radioactive hormone was secreted, but dibutyryl cyclic AMP, an agent that can mobilize stored parathormone, when added to the incubation mixtures enhanced radioactive parathormone secretion but only after 60 min, although it increased net hormone secretion as determined by radioimmunoassay to the same extent at all times studied. When the ionized calcium concentration of the medium was lowered, more radioactive hormone was secreted at all times but the effect was greatest on that hormone that was synthesized less than 60 min previously ; however, net hormone secretion in contrast to radioactive hormone was enhanced equally at all intervals. These data could mean that the refractoriness to secretion of parathormone 40-60 min of age was related to maturation of secretory container preparatory to storage. Low calcium (0.5 mM) stimulated hormone secretion up to fivefold compared to high calcium (3.0 mM) but did not affect synthesis of parathormone or proparathormne or conversion of the latter to hormone. During processing at least 70 percent of the intracellular parathormone was lost, presumably through proteolysis and this degradation was greater at high calcium. These data have been interpreted in light of the concept that two secretable pools of parathormone exist within the parathyroid. 相似文献
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Expression of N-linked sialyl Le(x) determinants and O-glycans in the carbohydrate moiety of human amniotic fluid transferrin during pregnancy 总被引:3,自引:0,他引:3
Transferrin, a glycoprotein involved in iron transport in body fluids, was
isolated from amniotic fluid of a hydramniospatient by sequential
anion-exchange chromatography and gel filtration. The N-glycans of human
amniotic fluid transferrin (hAFT) were enzymatically liberated by PNGase-F
digestion, isolated by gel filtration and fractionated by (high-pH)
anion-exchange chromatography. After alkaline borohydride treatment of
native hAFT, the released O-glycans were isolated by gel filtration and
fractionated by anion-exchange chroma-tography. Structure elucidation of 14
N- and 2 O-glycans was performed by 500 or 600 MHz1H-NMR spectroscopy.
Besides conventional N-glycans established earlier for human serum
transferrin (hST), new (alpha1-3)-fucosylated N- glycans were found,
representing sialyl Le(x) elements. Furthermore, as compared to hST, a
higher degree of (alpha1-6)-fucosylation and an increase in branching from
di- to triantennary compounds has been detected. The presence of O-glycans
is demonstrated for the first time in transferrin.
相似文献
110.