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91.
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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CLAIR. E. CALMET JOSIE LAMBOURDIRE JAWAD ABDELKRIM MICHEL PASCAL SARAH SAMADI 《Molecular ecology resources》2004,4(3):426-428
We developed eight polymorphic microsatellite loci to study the natural populations of Crocidura suaveolens, the lesser white‐toothed shrew. These loci are used in the study of insular populations of the French Atlantic coast where these shrews exhibit gigantism features that suggest an insular syndrome. These populations were threatened by the invasion of the rat Rattus norvegicus and the eradication of this alien pest in some islands was followed by a demographic expansion of the shrew populations. These first genetic results suggest that the shrew populations in the three studied islands are very differentiated. 相似文献
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Adhesion of phospholipid vesicles to Chinese hamster fibroblasts: Role of cell surface proteins 下载免费PDF全文
The adhesion of artificially generated lipid membrane vesicles to Chinese hamster V79 fibroblasts in suspension was used as a model system for studying membrane interactions. Below their gel-liquid crystalline phase transition temperature, vesicles comprised of dipalmitoyl lecithin (DPL) or dimyristoyl lecithin (DML) absorbed to the surfaces of EDTA- dissociated cells. These adherent vesicles could not be removed by repeated washings of the treated cells but could be released into the medium by treatment with trypsin. EM autoradiographic studies of cells treated with[(3)H]DML or [(3)H]DPL vesicles showed that most of the radioactive lipids were confined to the cell periphery. Scanning electron microscopy and fluorescence microscopy further confirmed the presence of adherent vesicles at the cell surface. Adhesion of DML or DPL vesicles to EDTA-dissociated cells modified the lactoperoxidase-catalyzed iodination pattern of the cell surface proteins; the inhibition of labeling of two proteins with an approximately 60,000- dalton mol wt was particularly evident. Incubation of cells wit h (3)H-lipid vesicles followed by sodium dodecyl sulfate (SDS)- polyacrylamide gel electrophoresis showed that some of the (3)H-lipid migrated preferentially with these approximately 60,000-mol wt proteins. Studies of the temperature dependence of vesicle uptake and subsequent release by trypsin showed that DML or DPL vesicle adhesion to EDTA- dissociated cells increased with decreasing temperatures. In contrast, cells trypsinized before incubation with vesicles showed practically no temperature dependence of vesicle uptake. These results suggest two pathways for adhesion of lipid vesicles to the cell surface-a temperature-sensitive one involving cell surface proteins, and a temperature-independent one. These findings are discussed in terms of current models for cell-cell interactions. 相似文献