
Determination of incompatibility group plasmids
kathryn
- 0
Determination of incompatibility group plasmids and copy number of the bla NDM-1 gene in carbapenem-resistant Klebsiella pneumoniae strains recovered from different hospitals in Kerman, Iran
Introduction. New Delhi metallo-β-lactamase (NDM)-producing Klebsiella pneumoniae has become a serious global health concern.
Hypothesis/Gap Statement. Due to the high genetic diversity among NDM-positive K. pneumoniae, we need further surveillance and studies to better understand the relationships between them. In addition, the coexistence of several plasmid replicon types in NDM-positive K. pneumoniae may affect the copy number of bla NDM, the MIC level to antibiotics, as well as increasing the chance of horizontal gene transfer.
Aim. The aim of this study was to determine incompatible plasmid groups and copy numbers of bla NDM, and to investigate the genetic relationship of 37 NDM-positive K. pneumoniae in Kerman, Iran.
Methodology. The bla NDM-1 gene was detected and confirmed by PCR-sequencing. The plasmid replicon types were determined by PCR-based replicon typing (PBRT) and the copy number of bla NDM-1 was determined by quantitaive real time-PCR (qPCR). Random amplified polymorphic DNA (RAPD)-PCR typing was used to detect genetic relationships between the strains.
Results. In this study, 10 different replicon types, including Frep [n=25 (67.5 %)], FIIAs [n=11 (29.7 %)], FIA [n=5 (13.5 %)], FIB [n=3 (8.1 %)], I1-Iγ [n=2 (5.4 %)], L/M [n=7 (18.9 %)], A/C [n=7 (18.9 %)], Y [n=3 (8.1 %)], P [n=1 (2.7 %)] and FIC [n=1 (2.7 %)] were reported. The copy numbers of the bla NDM-1 gene varied from 30.00 to 5.0×106 and no statistically significant correlation was observed between a rise of the MIC to imipenem and the copy numbers of bla NDM-1 (P>0.05). According to RAPD typing results, 35 strains were divided into five clusters, while two strains were non-typeable.Conclusion. The spread of NDM-1-producing K. pneumoniae strains that carry several plasmid replicon types increases the chance of horizontal transfer of antibiotic resistance genes in hospital settings. In this study, 10 different replicon types were identified. We could not find any relationship between the increase of MIC levels to imipenem and the copy numbers of bla NDM-1. Therefore, due to the identification of different replicon types in this study, the type and genetic characteristics of bla NDM-1-carrying plasmids, and other factors such as antibiotic selective pressure, probably affect the copy number of bla NDM-1 and change the MIC level to imipenem.

pOET8.VE1 transfer plasmid |
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200121 | Oxford Expression Technologies | 10 µg | EUR 409.2 |
pOET8.VE2 transfer plasmid |
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200122 | Oxford Expression Technologies | 10 µg | EUR 409.2 |
pOET8.VE3 transfer plasmid |
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200123 | Oxford Expression Technologies | 10 µg | EUR 409.2 |
pOET9 CMV transfer plasmid |
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200133 | Oxford Expression Technologies | 10 µg | EUR 280.24 |
pOET9 EF1α transfer plasmid |
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200131 | Oxford Expression Technologies | 10 µg | EUR 280.24 |
pOET9 CCAG transfer plasmid |
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pOET9 SV40 transfer plasmid |
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200134 | Oxford Expression Technologies | 10 µg | EUR 280.24 |
pOET2.1N/C_6xHis transfer plasmid |
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2001031 | Oxford Expression Technologies | 10 µg | EUR 208.32 |
pOET6 BacMAM transfer plasmid |
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200107 | Oxford Expression Technologies | 10 µg | EUR 208.32 |
TransferTip ES cell transfer |
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100µL Transfer Pipettes |
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TP100 | BioAssay Systems | N/A | EUR 15 |
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Description: Propane Transfer System |
Thermal Transfer Ribbon |
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WESTERN TRANSFER MODULE |
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WESTERN TRANSFER SYSTEM, |
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Rat cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Rat cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Pig cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Pig cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Goat cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Goat cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Goat cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Rabbit cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Rabbit cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Rabbit cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Human cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Human cholest erolester transfer protein/lipid transfer protein ELISA kit |
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E01C0732-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
Description: A sandwich ELISA for quantitative measurement of Human cholest erolester transfer protein/lipid transfer protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Human cholest erolester transfer protein/lipid transfer protein ELISA kit |
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E01C0732-96 | BlueGene | 1 plate of 96 wells | EUR 822 |
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Mouse cholest erolester transfer protein/lipid transfer protein ELISA kit |
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E03C0732-192T | BlueGene | 192 tests | EUR 1524 |
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Mouse cholest erolester transfer protein/lipid transfer protein ELISA kit |
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E03C0732-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
Description: A sandwich ELISA for quantitative measurement of Mouse cholest erolester transfer protein/lipid transfer protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Mouse cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Monkey cholest erolester transfer protein/lipid transfer protein ELISA kit |
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E09C0732-192T | BlueGene | 192 tests | EUR 1524 |
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Monkey cholest erolester transfer protein/lipid transfer protein ELISA kit |
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E09C0732-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
Description: A sandwich ELISA for quantitative measurement of Monkey cholest erolester transfer protein/lipid transfer protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Monkey cholest erolester transfer protein/lipid transfer protein ELISA kit |
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E09C0732-96 | BlueGene | 1 plate of 96 wells | EUR 822 |
Description: A sandwich ELISA for quantitative measurement of Monkey cholest erolester transfer protein/lipid transfer protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Plastic Transfer Pipets, graduated to 3ml, 500 pcs/pack |
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PTP-01 | IHC World | - | EUR 19.98 |
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PTP-02 | IHC World | - | EUR 19.98 |
CAPS Transfer Buffer 5X |
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21420016-1 | Bio-WORLD | 500 mL | EUR 60.54 |
Description: 3- (Cyclohexylamino) -1-propanesulfonic acid, N-Cyclohexyl-3-aminopropanesulfonic acid |
CAPS Transfer Buffer 5X |
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21420016-2 | Bio-WORLD | 1 L | EUR 91.52 |
Description: 3- (Cyclohexylamino) -1-propanesulfonic acid, N-Cyclohexyl-3-aminopropanesulfonic acid |
SPINE Puck Transfer Tool |
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M-SP-PTT-1 | MiTeGen | 1 UNIT | EUR 386 |
Description: SPINE Puck Transfer Tool |
CAPS Transfer Buffer 10X |
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20960009-1 | Bio-WORLD | 100 mL | EUR 26.06 |
Description: 3- (Cyclohexylamino) -1-propanesulfonic acid, N-Cyclohexyl-3-aminopropanesulfonic acid |
CAPS Transfer Buffer 10X |
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Description: 3- (Cyclohexylamino) -1-propanesulfonic acid, N-Cyclohexyl-3-aminopropanesulfonic acid |
CAPS Transfer Buffer 10X |
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20960009-3 | Bio-WORLD | 1 L | EUR 148.28 |
Description: 3- (Cyclohexylamino) -1-propanesulfonic acid, N-Cyclohexyl-3-aminopropanesulfonic acid |
Phospholipid transfer Antibody |
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20-abx110096 | Abbexa |
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Rapid Transfer Buffer 10X |
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10230000-1 | Bio-WORLD | 1 L | Ask for price |
Rapid Transfer Buffer 10X |
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Transfer Rack 1.5/2.0 mL |
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Guinea pig cholest erolester transfer protein/lipid transfer protein ELISA kit |
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E05C0732-192T | BlueGene | 192 tests | EUR 1524 |
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Guinea pig cholest erolester transfer protein/lipid transfer protein ELISA kit |
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E05C0732-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
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Guinea pig cholest erolester transfer protein/lipid transfer protein ELISA kit |
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Recombinant Bovine Glycolipid transfer protein Protein, His-SUMO, E.coli-10ug |
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QP6098-ec-10ug | EnQuireBio | 10ug | EUR 391.2 |
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804777 | Scientific Laboratory Supplies | EACH | EUR 45.72 |
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IB95010 | IBI Scientific | BLOT PAPER, 0.38MM - 10/PACK | EUR 38.22 |
SSRL Cassette Transfer Handle |
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M-CP-111-003 | MiTeGen | 1 UNIT | EUR 460 |
Description: SSRL Cassette Transfer Handle |
Human cholest erolester transfer protein/lipid transfer protein,CETP/LTP ELISA Kit |
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201-12-1516 | SunredBio | 96 tests | EUR 528 |
Description: A quantitative ELISA kit for measuring Human in samples from biological fluids. |
Human cholest erolester transfer protein/lipid transfer protein(CETP/LTP)ELISA Kit |
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Human cholest erolester transfer protein/lipid transfer protein(CETP/LTP)ELISA Kit |
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Human cholest erolester transfer protein/lipid transfer protein(CETP/LTP)ELISA Kit |
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Human cholest erolester transfer protein/lipid transfer protein,CETP/LTP ELISA Kit |
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Human cholest erolester transfer protein/lipid transfer protein,CETP/LTP ELISA Kit |
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Human cholest erolester transfer protein/lipid transfer protein,CETP/LTP ELISA Kit |
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Description: Cryo Transfer Face for SEM |
Cryo Transfer Face for TEM |
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Description: Cryo Transfer Face for TEM |
Alkaline transfer solution (10X) |
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10750000-1 | Bio-WORLD | 1 L | EUR 68.63 |
Phospholipid transfer Antibody (HRP) |
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20-abx108612 | Abbexa |
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Thermal Transfer Ribbon 1x2.7 OD |
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HIS3524 | Scientific Laboratory Supplies | EACH | EUR 244.8 |
Bis-Tris Transfer Buffer 20X |
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Recombinant Human Alpha-tocopherol transfer protein Protein, His-SUMO, E.coli-10ug |
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ARG1836 | Scientific Laboratory Supplies | PK250 | EUR 34.34 |
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Xpert Native Transfer Buffer(10X) |
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P9001-100 | GenDepot | 1L | EUR 165.6 |
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20810005-1 | Bio-WORLD | 1 Kit | EUR 128.98 |
Description: EzFastBlo |
Tris Glycine Transfer Buffer 10X |
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20960006-1 | Bio-WORLD | 1 L | EUR 66.99 |
Tris Glycine Transfer Buffer 10X |
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20960006-2 | Bio-WORLD | 4 L | EUR 173.72 |
Tris Glycine Transfer Buffer 10X |
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20960006-3 | Bio-WORLD | 500 mL | EUR 46.97 |
Phospholipid transfer Antibody (Biotin) |
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20-abx105774 | Abbexa |
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PAGE Gel Transfer Buffer, 10 x |
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GR103018 | Genorise Scientific | 1 L | EUR 79 |
Tris Glycine Transfer Buffer Packs |
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42020403-1 | Bio-WORLD | 40 PK (s) | EUR 109.86 |
ExoFectin Plasmid DNA-into-Exosome Kit (DNA transfection kit for exosome) |
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P402 | 101Bio | - | Ask for price |
Phospholipid transfer Polyclonal Antibody |
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42297-100ul | SAB | 100ul | EUR 399.6 |
FastBlot Transfer Buffer 5X, pH 11.0 |
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40320062-1 | Bio-WORLD | 500 mL | EUR 41.3 |
Semi Dry Blot Transfer Buffer 10X |
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21420067-1 | Bio-WORLD | 1 L | EUR 32.46 |
Semi Dry Blot Transfer Buffer 10X |
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21420067-2 | Bio-WORLD | 2 L | EUR 63.14 |
SuperBlot Western Transfer buffer 10X |
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DG-WEST | Bio Basic | 500ml | EUR 136.56 |
Human Transfer factor,TF ELISA Kit |
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201-12-1733 | SunredBio | 96 tests | EUR 528 |
Description: A quantitative ELISA kit for measuring Human in samples from biological fluids. |
Human Transfer factor,TF ELISA Kit |
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CN-04584H1 | ChemNorm | 96T | EUR 520.8 |
Human Transfer factor,TF ELISA Kit |
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CN-04584H2 | ChemNorm | 48T | EUR 340.8 |
Human Transfer factor(TF)ELISA Kit |
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GA-E1749HM-48T | GenAsia Biotech | 48T | EUR 346.8 |
Human Transfer factor(TF)ELISA Kit |
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GA-E1749HM-96T | GenAsia Biotech | 96T | EUR 559.2 |
Human Transfer factor(TF)ELISA Kit |
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QY-E00085 | Qayee Biotechnology | 96T | EUR 472.8 |
Human Transfer factor,TF ELISA Kit |
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YLA1849HU-48T | Shanghai YL Biotech | 48T | EUR 435 |
Human Transfer factor,TF ELISA Kit |
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YLA1849HU-96T | Shanghai YL Biotech | 96T | EUR 562.5 |
Human Transfer factor,TF ELISA Kit |
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SL1731Hu | Sunlong | 96 Tests | EUR 468 |
3 ml Transfer Pipets, 500 pcs/pack |
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30-0138 | IHC World | - | EUR 19 |
Bovine Glycolipid transfer protein (GLTP) |
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1-CSB-YP009539BO | Cusabio |
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Description: Recombinant Bovine Glycolipid transfer protein(GLTP) expressed in Yeast |
Bovine Glycolipid transfer protein (GLTP) |
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1-CSB-EP009539BO | Cusabio |
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Description: Recombinant Bovine Glycolipid transfer protein(GLTP) expressed in E.coli |
Transfer Pipette 23.0mL Extra Long 300mm |
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ARG1840 | Scientific Laboratory Supplies | PK100 | EUR 31.6 |
Transfer Pipette 6mL Extra Long 230mm |
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ARG1860 | Scientific Laboratory Supplies | PK500 | EUR 48.05 |
T-Pro Transfer Blotting buffer (10X) |
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JB08-H001 | T-Pro Biotechnology | 500ml/BT | EUR 162 |
Glycolipid Transfer Protein (GLTP) Protein |
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20-abx262570 | Abbexa |
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Human Phospholipid transfer protein (PLTP) |
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1-CSB-YP018212HU | Cusabio |
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Description: Recombinant Human Phospholipid transfer protein(PLTP) expressed in Yeast |
Human Phospholipid transfer protein (PLTP) |
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1-CSB-EP018212HU | Cusabio |
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Description: Recombinant Human Phospholipid transfer protein(PLTP) expressed in E.coli |
Glycolipid Transfer Protein (GLTP) Antibody |
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abx233495-100ug | Abbexa | 100 ug | EUR 610.8 |
Glycolipid Transfer Protein (GLTP) Antibody |
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20-abx112809 | Abbexa |
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Glycolipid Transfer Protein (GLTP) Antibody |
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20-abx128414 | Abbexa |
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Glycolipid Transfer Protein (GLTP) Antibody |
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20-abx303992 | Abbexa |
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Jumbo Transfer Pipette 10 None Sterile |
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F87928 | Scientific Laboratory Supplies | PK100 | EUR 24.05 |
Protein Stain Kit for Transfer Membrane |
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BSP025 | Bio Basic | 10Blots, 10prep | EUR 89.23 |
Phospholipid Transfer Protein (PLTP) Antibody |
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20-abx174059 | Abbexa |
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Phospholipid Transfer Protein (PLTP) Antibody |
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20-abx174060 | Abbexa |
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Phospholipid Transfer Protein (PLTP) Antibody |
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20-abx178016 | Abbexa |
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Phospholipid Transfer Protein (PLTP) Antibody |
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20-abx178017 | Abbexa |
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Phospholipid Transfer Protein (PLTP) Antibody |
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20-abx212015 | Abbexa |
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Phospholipid Transfer Protein (PLTP) Antibody |
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abx146426-100ug | Abbexa | 100 ug | EUR 469.2 |
Phospholipid Transfer Protein (PLTP) Antibody |
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abx033529-400ul | Abbexa | 400 ul | EUR 627.6 |
Phospholipid Transfer Protein (PLTP) Antibody |
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abx033529-80l | Abbexa | 80 µl | EUR 343.2 |
Phospholipid Transfer Protein (PLTP) Antibody |
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20-abx131266 | Abbexa |
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Phospholipid Transfer Protein (PLTP) Antibody |
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20-abx131267 | Abbexa |
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Phospholipid Transfer Protein (PLTP) Antibody |
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20-abx004303 | Abbexa |
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Phospholipid Transfer Protein (PLTP) Antibody |
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20-abx142148 | Abbexa |
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Phospholipid transfer protein Antibody / PLTP |
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RQ6393 | NSJ Bioreagents | 100 ug | EUR 356.15 |
Description: Phospholipid transfer protein (PLTP), also known as lipid transfer protein II is a protein that in humans is encoded by the PLTP gene. This gene is mapped to 20q13.12. The protein encoded by this gene is one of at least two lipid transfer proteins found in human plasma. The encoded protein transfers phospholipids from triglyceride-rich lipoproteins to high density lipoprotein (HDL). In addition to regulating the size of HDL particles, this protein may be involved in cholesterol metabolism. At least two transcript variants encoding different isoforms have been found for this gene. |
Recombinant Glycolipid Transfer Protein (GLTP) |
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4-RPC502Hu01 | Cloud-Clone |
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Description: Recombinant Human Glycolipid Transfer Protein expressed in: E.coli |
Recombinant Glycolipid Transfer Protein (GLTP) |
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RPU52857-100ug | Biomatik Corporation | 100ug | EUR 537.9 |
Recombinant Glycolipid Transfer Protein (GLTP) |
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RPU52857-1mg | Biomatik Corporation | 1mg | EUR 2380.3 |
Recombinant Glycolipid Transfer Protein (GLTP) |
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RPU52857-50ug | Biomatik Corporation | 50ug | EUR 431.2 |
Transfer Pipette 1.2mL General Purpose 64mm |
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ARG1834 | Scientific Laboratory Supplies | PK500 | EUR 35.64 |
Transfer Pipette 1.7mL General Purpose 87mm |
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ARG1838 | Scientific Laboratory Supplies | PK500 | EUR 35.64 |
Transfer Pipette 7.5mL Large Bulb Graduated |
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ARG1864 | Scientific Laboratory Supplies | PK500 | EUR 34.8 |
Rat Glycolipid Transfer Protein ELISA kit |
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E02G0192-192T | BlueGene | 192 tests | EUR 1524 |
Description: A competitive ELISA for quantitative measurement of Rat Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Rat Glycolipid Transfer Protein ELISA kit |
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E02G0192-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
Description: A competitive ELISA for quantitative measurement of Rat Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Rat Glycolipid Transfer Protein ELISA kit |
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E02G0192-96 | BlueGene | 1 plate of 96 wells | EUR 822 |
Description: A competitive ELISA for quantitative measurement of Rat Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Pig Glycolipid Transfer Protein ELISA kit |
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E07G0192-192T | BlueGene | 192 tests | EUR 1524 |
Description: A competitive ELISA for quantitative measurement of Porcine Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Pig Glycolipid Transfer Protein ELISA kit |
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E07G0192-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
Description: A competitive ELISA for quantitative measurement of Porcine Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Pig Glycolipid Transfer Protein ELISA kit |
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E07G0192-96 | BlueGene | 1 plate of 96 wells | EUR 822 |
Description: A competitive ELISA for quantitative measurement of Porcine Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Dog Glycolipid Transfer Protein ELISA kit |
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E08G0192-192T | BlueGene | 192 tests | EUR 1524 |
Description: A competitive ELISA for quantitative measurement of Canine Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Dog Glycolipid Transfer Protein ELISA kit |
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E08G0192-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
Description: A competitive ELISA for quantitative measurement of Canine Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Dog Glycolipid Transfer Protein ELISA kit |
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E08G0192-96 | BlueGene | 1 plate of 96 wells | EUR 822 |
Description: A competitive ELISA for quantitative measurement of Canine Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Goat Glycolipid Transfer Protein ELISA kit |
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E06G0192-192T | BlueGene | 192 tests | EUR 1524 |
Description: A competitive ELISA for quantitative measurement of Goat Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Goat Glycolipid Transfer Protein ELISA kit |
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E06G0192-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
Description: A competitive ELISA for quantitative measurement of Goat Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Goat Glycolipid Transfer Protein ELISA kit |
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E06G0192-96 | BlueGene | 1 plate of 96 wells | EUR 822 |
Description: A competitive ELISA for quantitative measurement of Goat Glycolipid Transfer Protein in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
Recombinant Phospholipid Transfer Protein (PLTP) |
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4-RPC728Mu01 | Cloud-Clone |
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Description: Recombinant Mouse Phospholipid Transfer Protein expressed in: E.coli |
Recombinant Phospholipid Transfer Protein (PLTP) |
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4-RPC728Ra01 | Cloud-Clone |
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Description: Recombinant Rat Phospholipid Transfer Protein expressed in: E.coli |
Recombinant Phospholipid Transfer Protein (PLTP) |
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RPU53605-100ug | Biomatik Corporation | 100ug | EUR 554.4 |
Plasmids shape the diverse accessory resistomes of Escherichia coli ST131
The human gut microbiome includes beneficial, commensal and pathogenic bacteria that possess antimicrobial resistance (AMR) genes and exchange these predominantly through conjugative plasmids. Escherichia coli is a significant component of the gastrointestinal microbiome and is typically non-pathogenic in this niche. In contrast, extra-intestinal pathogenic E. coli (ExPEC) including ST131 may occupy other environments like the urinary tract or bloodstream where they express genes enabling AMR and host cell adhesion like type 1 fimbriae.
The extent to which commensal E. coli and uropathogenic ExPEC ST131 share AMR genes remains understudied at a genomic level, and we examined this here using a preterm infant resistome. We found that individual ST131 had small differences in AMR gene content relative to a larger shared resistome. Comparisons with a range of plasmids common in ST131 showed that AMR gene composition was driven by conjugation, recombination and mobile genetic elements.
Plasmid pEK499 had extended regions in most ST131 Clade C isolates, and it had evidence of a co-evolutionary signal based on protein-level interactions with chromosomal gene products, as did pEK204 that had a type IV fimbrial pil operon. ST131 possessed extensive diversity of selective type 1, type IV, P and F17-like fimbriae genes that was highest in subclade C2. The structure and composition of AMR genes, plasmids and fimbriae vary widely in ST131 Clade C and this may mediate pathogenicity and infection outcomes.
Development of a simplified and inexpensive RNA depletion method for plasmid DNA purification using size selection magnetic beads (SSMBs)
Plasmid DNA (pDNA) isolation from bacterial cells is one of the most common and critical steps in molecular cloning and biomedical research. Almost all pDNA purification involves disruption of bacteria, removal of membrane lipids, proteins and genomic DNA, purification of pDNA from bulk lysate, and concentration of pDNA for downstream applications. While many liquid-phase and solid-phase pDNA purification methods are used, the final pDNA preparations are usually contaminated with varied degrees of host RNA, which cannot be completely digested by RNase A.
To develop a simple, cost-effective, and yet effective method for RNA depletion, we investigated whether commercially available size selection magnetic beads (SSMBs), such as Mag-Bind® TotalPure NGS Kit (or Mag-Bind), can completely deplete bacterial RNA in pDNA preparations. In this proof-of-principle study, we demonstrated that, compared with RNase A digestion and two commercial plasmid affinity purification kits, the SSMB method was highly efficient in depleting contaminating RNA from pDNA minipreps.
Gene transfection and bacterial colony formation assays revealed that pDNA purified from SSMB method had superior quality and integrity to pDNA samples cleaned up by RNase A digestion and/or commercial plasmid purification kits. We further demonstrated that the SSMB method completely depleted contaminating RNA in large-scale pDNA samples. Furthermore, the Mag-bind-based SSMB method costs only 5-10% of most commercial plasmid purification kits on a per sample basis. Thus, the reported SSMB method can be a valuable and inexpensive tool for the removal of bacterial RNA for routine pDNA preparations.
Emergence of plasmid-mediated tigecycline resistance tet(X4) gene in Escherichia coli isolated from poultry, food and the environment in South Asia
The recent emergence of mobile-tigecycline resistance tet(X) genes in human and animals in China seriously threats the clinical utility of tigecycline. Here we focused on the isolation and molecular characterization of plasmid-mediated tigecycline resistance tet(X4)-positive E. coli from different sources in Pakistan using MinION and Illumina sequencing. The tet(X4) gene was detected in four E. coli isolates from poultry, chicken meat, wild bird and the slaughterhouse wastewater in Pakistan. Co-existence of colistin resistance mcr-1 gene was also detected in three isolates.
The four isolates belonged to different sequence types and the tet(X4) gene was located on plasmids ranging from 12,331 bp to 113,610 bp belonging to IncFII and IncQ replicon types with two genetic contexts ISCR2-tet(X4)-abh-ISCR2-lysR-floR-virD2 and ΔISCR2-abh-tet(X4)-ISCR2-virD2-floR, respectively. In all the four E. coli strains, tet(X4) was transferable by conjugation to E. coli J53 host strain. In addition, three of four strains transferred tet(X4) to a wild-type carbapenem resistant E. coli strain. To our knowledge, this is the first report of the emergence of plasmid-mediated tet(X4) gene from Pakistan. The convergence of tigecycline and colistin resistance in South Asia is a serious threat to human health.