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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
104.
Yap CC Wisco D Kujala P Lasiecka ZM Cannon JT Chang MC Hirling H Klumperman J Winckler B 《The Journal of cell biology》2008,180(4):827-842
Correct targeting of proteins to axons and dendrites is crucial for neuronal function. We showed previously that axonal accumulation of the cell adhesion molecule L1/neuron-glia cell adhesion molecule (NgCAM) depends on endocytosis (Wisco, D., E.D. Anderson, M.C. Chang, C. Norden, T. Boiko, H. Folsch, and B. Winckler. 2003. J. Cell Biol. 162:1317-1328). Two endocytosis-dependent pathways to the axon have been proposed: transcytosis and selective retrieval/retention. We show here that axonal accumulation of L1/NgCAM occurs via nondegradative somatodendritic endosomes and subsequent anterograde axonal transport, which is consistent with transcytosis. Additionally, we identify the neuronal-specific endosomal protein NEEP21 (neuron-enriched endosomal protein of 21 kD) as a regulator of L1/NgCAM sorting in somatodendritic endosomes. Down-regulation of NEEP21 leads to missorting of L1/NgCAM to the somatodendritic surface as well as to lysosomes. Importantly, the axonal accumulation of endogenous L1 in young neurons is also sensitive to NEEP21 depletion. We propose that small endosomal carriers derived from somatodendritic recycling endosomes can serve to redistribute a distinct set of membrane proteins from dendrites to axons. 相似文献
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Wisco D Anderson ED Chang MC Norden C Boiko T Fölsch H Winckler B 《The Journal of cell biology》2003,162(7):1317-1328
Neuronal polarity is, at least in part, mediated by the differential sorting of membrane proteins to distinct domains, such as axons and somata/dendrites. We investigated the pathways underlying the subcellular targeting of NgCAM, a cell adhesion molecule residing on the axonal plasma membrane. Following transport of NgCAM kinetically, surprisingly we observed a transient appearance of NgCAM on the somatodendritic plasma membrane. Down-regulation of endocytosis resulted in loss of axonal accumulation of NgCAM, indicating that the axonal localization of NgCAM was dependent on endocytosis. Our data suggest the existence of a dendrite-to-axon transcytotic pathway to achieve axonal accumulation. NgCAM mutants with a point mutation in a crucial cytoplasmic tail motif (YRSL) are unable to access the transcytotic route. Instead, they were found to travel to the axon on a direct route. Therefore, our results suggest that multiple distinct pathways operate in hippocampal neurons to achieve axonal accumulation of membrane proteins. 相似文献
108.
Secretion and degradation of parathormone as a function of intracellular maturation of hormone pools 下载免费PDF全文
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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Marie JJ Huysman Cindy Martens Klaas Vandepoele Jeroen Gillard Edda Rayko Marc Heijde Chris Bowler Dirk Inzé Van de Yves Peer Lieven De Veylder Wim Vyverman 《Genome biology》2010,11(2):1-19