Synthetic control of plasmid replication enables target- and self-curing of vectors and expedites genome engineering of Pseudomonas putida.

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Synthetic control of plasmid replication enables target- and self-curing of vectors and expedites genome engineering of Pseudomonas putida.

Genome engineering of non-conventional microorganisms requires the event of devoted artificial biology instruments. Pseudomonas putida is a Gram-negative, non-pathogenic soil bacterium broadly used for metabolic engineering owing to its versatile metabolism and excessive ranges of tolerance to several types of stress. Genome enhancing of P. putida largely depends on homologous recombination occasions, assisted by helper plasmid-based expression of genes encoding DNA modifying enzymes. Plasmid curing from chosen isolates is probably the most tedious and time-consuming step of this process, and implementing generally used strategies to this finish in P. putida (e.g. temperature-sensitive replicons) is usually impractical.

To deal with this difficulty, we have now developed a toolbox for each target- and self-curing of plasmid DNA in Pseudomonas species. Our technique allows plasmid-curing in a easy cultivation step by combining in vivo digestion of vectors by the I-SceI homing nuclease with artificial management of plasmid replication, triggered by the addition of an inexpensive chemical inducer (3-methylbenzoate) to the medium. The system shows an effectivity of vector curing >90% and the screening of plasmid-free clones is drastically facilitated by means of fluorescent markers that may be chosen in accordance with the appliance supposed.

Moreover, fast genome engineering of P. putida utilizing self-curing plasmids is demonstrated via genome discount of the platform pressure EM42 by eliminating all genes encoding β-lactamases, the catabolic ben gene cluster, and the pyoverdine synthesis equipment. Physiological characterization of the ensuing streamlined pressure, P. putida SEM10, revealed advantageous options that could possibly be exploited for metabolic engineering.

The plasmid vectors, pBS2ndd and pBS3ndd, for versatile cloning with low background in Escherichia coli.

For many years, various plasmid vectors have been repeatedly developed for molecular cloning of DNA fragment within the bacterial host cell Escherichia coli. Even with deliberate performances in vector preparation, the cloning approaches nonetheless face inevitable background colonies, or false optimistic clones, which may be arisen from intact or self-ligated plasmid molecules. To help in such drawback, two plasmids, pBS2ndd and pBS3ndd, which proof against ampicillin and kanamycin respectively, had been developed on this examine as extra advantageous cloning vector. The plasmids carry ndd, a deadly gene from bacteriophage T4 coding for nucleoid disruption protein that binds to the host chromosome and progressively kill the cell.

The lethal toxicity of Ndd inhibits host cells that receive intact or ndd-religated vector from rising, which leads to low background and dramatically reduces the trouble for choice of recombinants. Furthermore, their equivalent a number of cloning web site was designed to help varied cloning methods. Digestion of plasmids with XcmI permits for in vitro T/A ligation, whereas with EcoRV permits blunt-end ligation, with functionality of blue-white colony screening. In vivo homologous recombination cloning can be utilizable by amplification of insert fragments utilizing primers containing homology arms and transformation into succesful E. coli strains.

To reveal their benefits, the plasmids had been used to clone PCR product samples for DNA sequencing with low-background and versatile cloning methods. Such fast and cost-effective cloning procedures are additionally proposed right here. Lastly, the cloning for protein expression with blue-white choice was additionally potential utilizing egfp as a mannequin regulated by lac and T7 promoters on the plasmid or different build-in promoters with the insert.

Synthetic control of plasmid replication enables target- and self-curing of vectors and expedites genome engineering of Pseudomonas putida.

Building of a plasmid vector containing epidermal development issue receptor and C-Jun shRNA.

The target of this examine was to assemble a plasmid vector for EGFR-hm-1 and C-Junh-825 small interfering RNA (siRNA). EGFR-hm-1 and C-Jun-hm-825 oligonucleotide fragments had been synthesized and quick hairpin RNA (shRNA) had been amplified by PCR. Plasmids had been remoted from E. coli TOP10 bacterium by restriction enzyme digestion utilizing pst1 and BamH1 and oligonucleotide fragments had been cloned into the pSilencer plasmid containing the U6 promoter.

Recombinant clones had been generated by remodeling JM109 competent cells with plasmid vectors containing shRNA molecules. 58 base-paired EGFR-hm-1 and 59 base-paired C-Jun-hm-825 oligonucleotide fragments had been remoted. The fragments had been 100% homologous with human sequences out there on GenBank. The recombinant pSilencer1.zero vector containing a 58-bp EGFR-hm-1 and 59-bp C-Jun-hm-825 fragment was constructed. These vectors have the potential for use as remedy to fight pores and skin photoaging underneath UV publicity.

Characterization of a cryptic plasmid remoted from Lactobacillus casei CP002616 and development of shuttle vectors based mostly on its replicon.

The cryptic plasmid pLC2W was remoted from Lactobacillus casei CP002616. Nucleotide sequence evaluation revealed that four putative open studying frames (ORF) had been answerable for DNA replication. 4 Escherichia coli-Lactobacillus shuttle vectors had been constructed utilizing completely different lengths of the pLC2W replicon to establish the shortest practical replicon. The size of the pLC2W replicon didn’t have an effect on the soundness of the plasmids. Inexperienced fluorescent protein (GFP) as a reporter was expressed efficiently in a number of lactobacilli utilizing our constructed vectors.

The outcomes urged that the expression vectors pUE-F0GFP and pUE-F1GFP are potential molecular instruments for heterologous gene cloning and expression in lactobacilli. Furthermore, 2 plasmid-curing strategies had been used to remove pLC2W from L. casei. We detected no distinction between L. casei CP002616 and L. casei CP002616 pLC2WΔ-IC (mutant pressure cured by plasmid incompatibility technique) in manufacturing of exopolysaccharide (EPS) or acid.

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MBS-K060 0.5mg
EUR 3028.1
Description: Leukocyte surface antigen CD47 is also known as Antigenic surface determinant protein OA3, Integrin-associated protein (IAP) and Protein MER6. CD47 contains 1 Ig-like V-type (immunoglobulin-like) domain. CD47 is very broadly distributed on normal adult tissues. CD47 has a role in both cell adhesion by acting as an adhesion receptor for THBS1 on platelets, and in the modulation of integrins and plays an important role in memory formation and synaptic plasticity in the hippocampus by similarity. CD47 is the receptor for SIRPA, binding to which prevents maturation of immature dendritic cells and inhibits cytokine production by mature dendritic cells. CD47 Interaction with SIRPG mediates cell-cell adhesion, enhances superantigen-dependent T-cell-mediated proliferation and costimulates T-cell activation.

SARS-CoV-2 Spike S1-coupled Magnetic Beads
MBS-K001 2mg
EUR 2707.1
Description: SARS-CoV-2 Spike S1 protein is expressed from human 293 cells (HEK293). It contains AA Val 16 - Arg 685(Accession # QHD43416.1).

SARS-CoV-2 Spike S2-coupled Magnetic Beads
MBS-K018 2mg
EUR 2707.1
Description: SARS-CoV-2 Spike S2 Protein is expressed from human 293 cells (HEK293). It contains AA Ser 686 - Pro 1213 (Accession # QHD43416.1 (F817P, A892P, A899P, A942P,K986P,V987P)).

SARS-CoV-2 Spike RBD-coupled Magnetic Beads
MBS-K002 2mg
EUR 2707.1
Description: SARS-CoV-2 Spike RBD protein is expressed from human 293 cells (HEK293). It contains AA Arg 319 - Phe 541(Accession # QHD43416.1).

SARS-CoV-2 Spike NTD-coupled Magnetic Beads
MBS-K019 2mg
EUR 2707.1
Description: SARS-CoV-2 Spike NTD Protein is expressed from human 293 cells (HEK293). It contains AA Val 16 - Phe 318(Accession # QHD43416.1).

Human CD3E & CD3D Heterodimer-coupled Magnetic Beads
MBS-K003 2mg
EUR 2707.1
Description: Human CD3E & CD3D Heterodimer protein is expressed from human 293 cells (HEK293).It contains AA Asp 23 - Asp 126 (CD3E) & Phe 22 - Ala 105 (CD3D) (Accession # P07766-1(CD3E) & P04234-1(CD3D)).

Mouse monoclonal E2-Crimson antibody, Magnetic beads
TA183040 1 ml Ask for price

Magnetic Beads-conjugated Rabbit IgG isotype control
AC047 100μL
EUR 87.75
Description: None

SARS-CoV-2 Spike Trimer-coupled Magnetic Beads
MBS-K015 2mg
EUR 4312.1
Description: SARS-CoV-2 Spike Trimer protein is expressed from human 293 cells (HEK293). It contains AA Val 16 - Pro 1213(Accession # QHD43416.1 (R683A, R685A)).

Mouse monoclonal DsRed-Monomer antibody, Magnetic beads
TA183039 1 ml Ask for price

Magnetic Beads-Based Virus Genomic Nucleic Acid Kit
abx460018-1096tests 10 × 96 tests Ask for price

Magnetic Beads-Based Virus Genomic Nucleic Acid Kit
abx460018-596tests 5 × 96 tests Ask for price

Magnetic Beads-Based Virus Genomic Nucleic Acid Kit
abx460018-96tests 96 tests
EUR 437.5

Mouse monoclonal DsRed-Express2 antibody, Magnetic beads
TA183029 1 ml Ask for price

Mouse monoclonal mOrange/mOrange2 antibody, Magnetic beads
TA183035 1 ml Ask for price

Magnetic Beads-conjugated Mouse anti DDDDK-Tag mAb
AE037 50 ul
EUR 357.5

B2M mouse monoclonal antibody,clone 1A9, Magnetic Beads
TA802776JM 1 ml Ask for price

Magnetic Beads-conjugated Rabbit anti DDDDK-Tag mAb
AE097 1000μL
EUR 84.5

HA mouse monoclonal antibody, clone CB051, Magnetic Beads
TA183070 1 ml Ask for price

Mouse monoclonal mRaspberry/AmCyan1 antibody, Magnetic beads
TA183031 1 ml Ask for price

Protein A-coupled Magnetic Beads (recommended for MPCLIA)
MPC-A001 10mg
EUR 866.7
Description: The Protein A-coupled Magnetic Beads are 2.8 µm superparamagnetic particles covalently coupled to a highly pure form of protein A. The beads can be used to capture the antibodies in Chemiluminescence procedures.The Protein A is a 42 kDa surface protein, It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family.The Anti-Protein A-coupled Magnetic Beads is easy to capture the antibodies, and the bounded antibody have no activity lost, this ready to use products could greatly save your protein coupling time and hassle, and help us get the best performance and highly reproducible results.

Protein G-coupled Magnetic Beads (recommended for MPCLIA)
MPC-A002 10mg
EUR 866.7
Description: The Protein G-coupled Magnetic Beads are 2.8 µm superparamagnetic particles covalently coupled to a highly pure form of protein G. The beads can be used to capture the antibodies in Chemiluminescence procedures.The Protein G binds the heavy chain within the Fc region of most immunoglobulins but not bind dog immunoglobulins.The Protein G-coupled Magnetic Beads is easy to capture the most antibodies, and the bounded antibody have no activity lost, this ready to use products could greatly save your protein coupling time and hassle, and help us get the best performance and highly reproducible results.

Protein L-coupled Magnetic Beads (recommended for MPCLIA)
MPC-A008 10mg
EUR 866.7
Description: The Protein L-coupled Magnetic Beads are 2.8 µm superparamagnetic particles covalently coupled to a highly pure form of Protein L. The beads can be used to capture the antibodies in Chemiluminescence procedures. The Protein L is a 45 kDa surface protein, and can bind almost all antibody types, including IgG, IgM, IgA, IgE and IgD. It binds to antibodies with appropriate Kappa light chains such as VkI, VkIII and VkIV, whereas no binding occurred with antibodies of the VkII or with any Lambda light chains. The Protein L-coupled Magnetic Beads is easy to capture the most antibodies, and the bounded antibody have no activity lost, this ready to use products could greatly save your protein coupling time and hassle, and help us get the best performance and highly reproducible results.

Mouse monoclonal AcGFP1/PS-CFP2 antibody, Magnetic beads
TA183028 1 ml Ask for price

resDetect™ resDNA Sample Preparation Kit (Magnetic Beads)
OPA-R005 50preps
EUR 310.3
Description: DNA extraction is a critical step in many molecular biology techniques, such as PCR, sequencing, and cloning, as it is necessary to obtain pure and intact DNA that can be used in downstream applications. DNA extraction kits can simplify and standardize the DNA extraction process, allowing researchers to obtain high-quality DNA from a wide range of samples with reproducible results.

SARS-CoV-2 Spike RBD (B.1.351) Coupled Magnetic Beads
MBS-K032 2mg
EUR 2707.1
Description: SARS-CoV-2 Spike RBD (B.1.351) Protein is expressed from human 293 cells (HEK293). It contains AA Arg 319 - Lys 537(Accession # QHD43416.1 (K417N,E484K,N501Y)).

SARS-CoV-2 Spike RBD (P.1) Coupled Magnetic Beads
MBS-K033 2mg
EUR 2707.1
Description: SARS-CoV-2 Spike RBD (P.1) Protein is expressed from human 293 cells (HEK293). It contains AA Arg 319 - Lys 537(Accession # QHD43416.1 (K417T,E484K,N501Y)).

SARS-CoV-2 Spike RBD (B.1.1.7) Coupled Magnetic Beads
MBS-K034 2mg
EUR 2707.1
Description: SARS-CoV-2 Spike RBD (B.1.1.7) Protein is expressed from human 293 cells (HEK293). It contains AA Arg 319 - Lys 537(Accession # QHD43416.1(N501Y)).

SARS-CoV-2 (Delta) Spike RBD-coupled Magnetic Beads
MBS-K037 2mg
EUR 2707.1
Description: SARS-CoV-2 (Delta) Spike RBD Protein is expressed from human 293 cells (HEK293). It contains AA Arg 319 - Lys 537(Accession # QHD43416.1 (L452R, T478K)).

SARS-CoV-2 Spike Trimer (D614G)-coupled Magnetic Beads
MBS-K016 2mg
EUR 4312.1
Description: SARS-CoV-2 Spike Trimer (D614G) protein is expressed from human 293 cells (HEK293). It contains AA Val 16 - Pro 1213(Accession # QHD43416.1 (D614G, R683A, R685A)).

Universal IP/Co-IP Toolkit (Magnetic Beads/Anti-Mouse)
KTI1010-EN-20T 20 T
EUR 139
Description: Antibody coupling magnetic beads, suitable for IP/ CO-IP experiment, protein purification.

Mouse monoclonal Timer antibody, clone OTI3C9, Magnetic beads
TA183026 1 ml Ask for price

SARS-CoV-2 (Omicron) Spike RBD-coupled Magnetic Beads
MBS-K043 2mg
EUR 2707.1
Description: SARS-CoV-2 (Omicron) Spike RBD Protein is expressed from human 293 cells (HEK293). It contains AA Arg 319 - Lys 537(Accession # QHD43416.1 (G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493K, G496S, Q498R, N501Y, Y505H)).

Universal IP/Co-IP Toolkit (Magnetic Beads/Anti-Rabbit)
KTI1020-EN-20T 20 T
EUR 139
Description: Antibody coupling magnetic beads, suitable for IP/ CO-IP experiment, protein purification.

SARS-CoV-2 Nucleocapsid Protein-coupled Magnetic Beads
MBS-K017 2mg
EUR 2707.1
Description: SARS-CoV-2 Nucleocapsid Protein is expressed from human 293 cells (HEK293). It contains AA Met 1 - Ala 419(Accession # QHO62115.1).

SARS-CoV-2 Spike Trimer (B.1.1.7) Coupled Magnetic Beads
MBS-K029 2mg
EUR 2707.1
Description: SARS-CoV-2 Spike Trimer (B.1.1.7) Protein is expressed from human 293 cells (HEK293). It contains AA Val 16 - Pro 1213(Accession # QHD43416.1 (HV69-70del/Y144del//N501Y/A570D/D614G/P681H/T716I/S982A/D1118H/R683A/R685A/F817P/A892P/A899P/A942P/K986P/V987P)).

SARS-CoV-2 Spike Trimer (B.1.351) Coupled Magnetic Beads
MBS-K030 2mg
EUR 2707.1
Description: SARS-CoV-2 Spike Trimer (B.1.351) Protein is expressed from human 293 cells (HEK293). It contains AA Val 16 - Pro 1213(Accession # QHD43416.1 (L18F, D80A, D215G, 242-244del, R246I, K417N, E484K, N501Y, D614G, A701V, R683A, R685A,F817P, A892P, A899P, A942P, K986P, V987P)).

SARS-CoV-2 Spike Trimer (P.1) Coupled Magnetic Beads
MBS-K031 2mg
EUR 2707.1
Description: SARS-CoV-2 Spike Trimer (P.1) Protein is expressed from human 293 cells (HEK293). It contains AA Val 16 - Pro 1213(Accession # QHD43416.1 (L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, T1027I, V1176F, R683A, R685A,F817P, A892P, A899P, A942P, K986P, V987P)).

SARS-CoV-2 (Delta) Spike Trimer-coupled Magnetic Beads
MBS-K038 2mg
EUR 4312.1
Description: SARS-CoV-2 (Delta) Spike Trimer Protein is expressed from human 293 cells (HEK293). It contains AA Arg 319 - Lys 537(Accession # QHD43416.1 (T19R, G142D, EF156-157del, R158G, L452R, T478K, D614G, P681R, D950N, R683A, R685A, F817P, A892P, A899P, A942P, K986P, V987P)).

Mouse monoclonal ZsGreen1 antibody, clone OTI2C2, Magnetic beads
TA183023 1 ml Ask for price

Anti-Mouse IgG-coupled Magnetic Beads (recommended for MPCLIA)
MPC-A003 10mg
EUR 1305.4
Description: The Anti-Mouse IgG-coupled Magnetic Beads are 2.8 µm superparamagnetic particles covalently coupled to a highly affinity polyclonal goat anti-mouse IgG antibody. The beads can be used to capture the mouse IgG in Chemiluminescence procedures.The antibody reacts with the Fc or Fab portion of mouse IgG. No antibody was detected against mouse IgM or IgA, or against non-immunoglobulin serum proteins. The antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with bovine serum proteins, but it may cross-react with immunoglobulins from other species.The Anti-Mouse IgG-coupled Magnetic Beads is easy to capture the mouse IgG, and the bounded protein have no activity lost, this ready to use products could greatly save your protein coupling time and hassle, and help us get the best performance and highly reproducible results.

Anti-Human IgG-coupled Magnetic Beads (recommended for MPCLIA)
MPC-A004 10mg
EUR 984.4
Description: The Anti-Human IgG-coupled Magnetic Beads are 2.8 µm superparamagnetic particles covalently coupled to a highly affinity Monoclonal mouse anti-human IgG antibody. The beads can be used to capture the human IgG in Chemiluminescence procedures.The antibody reacts with the Fc portion of human IgG heavy chain but not with the Fab portion of human IgG. No antibody was detected against human IgM or IgA, or against non-immunoglobulin serum proteins. The antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with mouse, cynomolgus and bovine serum proteins, but it is not sure if the product cross-react with immunoglobulins from other species.The Anti-Human IgG-coupled Magnetic Beads is easy to capture the human IgG, and the bounded antibody have no activity lost, this ready to use products could greatly save your protein coupling time and hassle, and help us get the best performance and highly reproducible results.

Anti-Human IgM-coupled Magnetic Beads (recommended for MPCLIA)
MPC-A010 100mg
EUR 107
Description: The Anti-Human IgM-coupled Magnetic Beads are 2.8 µm superparamagnetic particles covalently coupled to a highly affinity Monoclonal anti-human IgM antibody. The beads can be used to capture the Human IgM in Chemiluminescence procedures. It has no-corss-reactivity with Human IgG, IgA or IgE. The Anti-Human IgM-coupled Magnetic Beads is easy to capture the human IgM, and the bounded antibody have no activity lost, this ready to use products could greatly save your protein coupling time and hassle, and help us get the best performance and highly reproducible results.

Mouse monoclonal mGFP/mCFP/mYFP/KillerRed antibody, Magnetic beads
TA183038 1 ml Ask for price

DiagNano™ PEG Magnetic Quantum Dot Beads, 655 nm Emission Peak
DNQ-CQ012 150 ul
EUR 960

DiagNano™ CD4 Magnetic Quantum Dot Beads, 655 nm Emission Peak
DNQ-CQ013 150 ul
EUR 960

DiagNano™ PEG Magnetic Quantum Dot Beads, 610 nm Emission Peak
DNQ-CQ015 150 ul
EUR 960

DiagNano™ CD3 Magnetic Quantum Dot Beads, 655 nm Emission Peak
DNQ-CQ001 20 tests
EUR 1160

DiagNano™ CD4 Magnetic Quantum Dot Beads, 610 nm Emission Peak
DNQ-CQ002 20 tests
EUR 1160

Nonetheless, EPS and acid manufacturing had been each diminished in L. casei CP002616 pLC2WΔ-HT (mutant pressure cured by high-temperature warmth remedy technique), demonstrating a distinction between these 2 curing strategies. Sequence evaluation of pLC2W and plasmid curing information recommend that plasmid pLC2W shouldn’t be concerned in EPS synthesis.

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