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Escherichia coli dh10b

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Escherichia coli DH10B is a bacterial strain commonly used in molecular biology laboratories. It is a genetically engineered derivative of the E. coli K-12 strain, designed to enhance the stability and transformation efficiency of DNA plasmids. The strain features a set of genetic modifications that improve the strain's capacity for cloning and recombinant DNA work.

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Lab products found in correlation

Escherichia coli bl21, by New England Biolabs (2 mentions) Escherichia coli hb101, by Promega (2 mentions) Tobramycin, by Merck Group (1 mentions) Paromomycin, by Merck Group (1 mentions) Glass bead, by Merck Group (1 mentions) Neomycin, by Merck Group (1 mentions) Amikacin, by Merck Group (1 mentions) Kanamycin b, by Merck Group (1 mentions) 2 mercaptoethanol, by Merck Group (1 mentions) Hplc grade acetonitrile, by Avantor (1 mentions) Yeast extract, by BD (1 mentions) Malt extract, by BD (1 mentions)

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Escherichia coli (4 citations) DH10B (3 citations) Escherichia coli K-12 DH10B (3 citations)

5 protocols using escherichia coli dh10b

1

Antibiotics and Nucleoside Precursor Preparation

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EXAMPLE 1

Preparation of Materials

Standard kanamycin, kanamycin B, neomycin, paromomycin, tobramycin, and amikacin were purchased from Sigma (USA). 2-mercaptoethanol, phenylmethyl-sulfonyl fluoride (PMSF), phenol/chloroform/isoamyl alcohol (25:24:1), uridine 5′-diphospho-D-glucose (UDP-Glc), and glass beads (150 to 212 μm) were also purchased from Sigma. Heptafluorobutyric acid (HFBA) was obtained from Fluka, and HPLC-grade acetonitrile, methanol, and water were obtained from J. T. Baker. 2-deoxystreptamine (2-DOS, compound 1) and UDP-2-N-acetyl-D-glucosamine (UDP-GlcNAc) were purchased from GeneChem (Republic of Korea). Cation solid-phase exchanger (OASIS MXC SPE, 3 mL/60 mg) and vacuum manifold were purchased from Waters. The culture medium components, soybean meal, yeast extract, and malt extract were acquired from BD Science (USA).

Paromamine and neamine were prepared from paromomycin and neomycin, respectively, by methanolysis. UDP-2-D-glucosamine (UDP-GlcN) was prepared by enzymatic reaction of UDP-D-glucose pyrophosphorylase with glucosamine-1-phosphate and uridine 5′-triphosphate (UTP). Escherichia coli DH10B and plasmid Litmus 28 (New England Biolabs) were used for routine subcloning. High-copy number E. coli-Streptomyces shuttle vector pSE34 containing the strong ermE* promoter plus a thiostrepton resistance marker was used as an expression plasmid.

US10337044B2. Kanamycin compound, kanamycin-producing Streptomyces species bacterium, and method of producing kanamycin (2019-07-02). iNtRON Biotechnology, Inc. [KR]. Inventors: Yeo Joon Yoon [KR], Sung Ryeol Park [KR], Je Won Park [KR], Jae Kyung Sohng [KR].
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2

Campylobacter jejuni Genetic Derivatives

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Campylobacter jejuni 11168 and its derivatives—C. jejunicmeD::cat, C. jejunicmeD::cat wtcmeABC, and C. jejunicmeD::cat g0cmeABC—were used in this study (Table 2). C. jejuni 11168H was grown on Columbia-based agar (CBA) or Muller-Hinton-based agar supplemented with 5% horse blood according to the manufacturer’s instructions. Strains were grown at 37°C in a variable atmospheric incubator (VAIN) cabinet (Don Whitely, UK) maintaining the following microaerophilic conditions: 85% nitrogen, 5% oxygen, and 10% carbon dioxide. All of the cloning experiments were performed in Escherichia coli DH10B (New England Biolabs). E. coli DH10B was used in the expression of CmeA and the cloning and expression of CmeC, whereas gCmeA was expressed in E. coli SBD1. E. coli strains were grown on either Luria-Bertani (LB) broth or LB agar, and antibiotics were added when necessary.
Abouelhadid S., Raynes J., Bui T., Cuccui J, & Wren B.W. (2020). Characterization of Posttranslationally Modified Multidrug Efflux Pumps Reveals an Unexpected Link between Glycosylation and Antimicrobial Resistance. mBio, 11(6), e02604-20.
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3

Escherichia coli Strains for Plasmid Preparation

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Escherichia coli HB101 (Promega) was used as the recipient strain for preparation of HIV-1 proviral vectors, whereas all of the other plasmid vectors were prepared from Escherichia coli DH10B (NEB). GST-fusion proteins were prepared from Escherichia coli BL21 (NEB). All these bacteria were cultured in LB broth in a shaking incubator at 37°C.
Chai Q., Li S., Collins M.K., Li R., Ahmad I., Johnson S.F., Frabutt D.A., Yang Z., Shen X., Sun L., Hu J., Hultquist J.F., Peterlin B.M, & Zheng Y.H. (2021). HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity. Cell reports, 36(6), 109514.
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4

Escherichia coli Strains for Plasmid Preparation

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Escherichia coli HB101 (Promega) was used as the recipient strain for preparation of HIV-1 proviral vectors, whereas all of the other plasmid vectors were prepared from Escherichia coli DH10B (NEB). GST-fusion proteins were prepared from Escherichia coli BL21 (NEB). All these bacteria were cultured in LB broth in a shaking incubator at 37°C.
Chai Q., Li S., Collins M.K., Li R., Ahmad I., Johnson S.F., Frabutt D.A., Yang Z., Shen X., Sun L., Hu J., Hultquist J.F., Peterlin B.M, & Zheng Y.H. (2021). HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity. Cell reports, 36(6), 109514.
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5

Cloning and Evolution Protocols in E. coli

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Escherichia coli DH10B (New England BioLabs, Ipswich, MA, USA) was used for all routine cloning and directed evolution. All biosensor systems were characterized in E. coli DH10B. LB-Miller (LB) media (BD, Franklin Lakes, NJ, USA) was used for routine cloning, fluorescence assays, directed evolution, and orthogonality assays unless specifically noted. LB + 1.5% agar (BD, Franklin Lakes, NJ, USA) plates were used for routine cloning and directed evolution. The plasmids described in this work were constructed using Gibson assembly, golden gate assembly, and standard molecular biology techniques. Synthetic genes, obtained as gBlocks, and primers were purchased from IDT. Relevant plasmid sequences are provided in Supplementary Table 4 and are available by request from the corresponding authors.
d'Oelsnitz S., Nguyen V., Alper H.S, & Ellington A.D. (2022). Evolving a Generalist Biosensor for Bicyclic Monoterpenes. ACS synthetic biology, 11(1).
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