Ecori

Manufactured by GenScript

Sourced in China

EcoRI is a type II restriction enzyme isolated from the bacterium Escherichia coli. It recognizes and cleaves the palindromic DNA sequence 5'-GAATTC-3' and its reverse complement 5'-CTTAAG-3'. EcoRI is widely used in molecular biology applications, such as DNA cloning and analysis.

Automatically generated - may contain errors

Lab products found in correlation

Hindiii, by GenScript (2 mentions) T4 dna ligase, by New England Biolabs (1 mentions) Ecori, by New England Biolabs (1 mentions) Phanta max super fidelity dna polymerase, by Vazyme (1 mentions) Pcdh cmv mcs ef1 puro, by System Biosciences (1 mentions) Bamhi, by GenScript (1 mentions) Superdex 200, by GE Healthcare (1 mentions) Complete his tag purification resin, by Roche (1 mentions) Superpose 6 10 300 gl, by GE Healthcare (1 mentions)

Spelling variants of this product

NdeI and EcoRI cloning sites (2 citations)

4 protocols using ecori

Cloning and Sequencing of slncRNA Constructs

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All sequences encoding for the in vitro slncRNAs (i.e., PP7-3x, PP7-4x, PP7-3x/MS2-3x, PP7-4x/MS2-4x, PP7-8x and PP7-14x/MS2-15x. Sequences appear in Supplementary Data 1) were ordered from Integrated DNA Technologies (IDT), (Coralville, Iowa) as gBlock gene fragments downstream to a T7 promoter and flanked by EcoRI (New England Biolabs (NEB),Ipswich, MA, #R3101L) restriction sites on both sides. gBlocks were cloned into a high-copy plasmid containing an Ampicillin resistance gene and verified using Sanger sequencing.
The 5Qβ/4PP7 slncRNA sequence was ordered from GenScript, Inc. (Piscataway, NJ), as part of a puc57 plasmid, flanked by EcoRI and HindIII (NEB, #R3104L) restriction sites. pBAC-lacZ backbone plasmid was obtained from Addgene (plasmid #13422). Both insert and vector were digested using the said restriction enzymes and ligated to form a circular plasmid using T4 DNA ligase (NEB, #M0202L). Sequence was verified by sanger sequencing.

Granik N., Katz N., Willinger O., Goldberg S, & Amit R. (2022). Formation of synthetic RNA protein granules using engineered phage-coat-protein -RNA complexes. Nature Communications, 13, 6811.

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Cloning and Expression of H-Fe Fusion Protein

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After codon preference optimization, the PPRV H protein (GenBank: AJE30413.1) and H. pylori ferritin (GenBank: NP_223316, 5-167aa, N19Q) sequences were synthesized using GenScript (Nanjing, China) with BamHI and EcoRI restriction sites at the upstream and downstream, respectively. The H protein sequence was fused to the N-terminal of ferritin by a linker (Ser-Gly-Gly). The H-Fe and H fragments were amplified via fusion PCR (Phanta Max Super-Fidelity DNA Polymerase was purchased from Vazyme), and then the H-Fe and H protein sequences were inserted into the pet28a vector by BamHI/EcoRI (FastDigest enzymes BamHI and EcoRI were purchased from Thermo Scientific) digestion to construct the vectors pET-28a-H-Fe and pET-28a-H.

Li D., Song H., Li J., Liu X., Gao X., Wu T., Zhang Z, & Li Y. (2022). Expression and Evaluation of a Novel PPRV Nanoparticle Antigen Based on Ferritin Self-Assembling Technology. Pharmaceutics, 14(9), 1902.

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YWHAE-NUTM2 Fusion Construct Generation

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A YWHAE–NUTM2–FLAG fusion cDNA containing BamHI (YWHAE sequence) and EcoRI (FLAG sequence) restriction sites was synthesized (GenScript) per the YWHAE–NUTM2 fusion transcript sequence in ESS1 and cloned in a pUC57 vector. The fusion gene sequence was validated by sequencing. It was further subcloned in pCDNA3(+) by EcoRI and BamHI (GenScript). YWHAE–NUTM2–FLAG was then subcloned into lentiviral vector pCDH-CMV-MCS-EF1-Puro (System Biosciences) by Nhe1 & Not1 (New England Biolabs). Construct integrity was verified by Sanger sequencing.

Ou W.B., Lundberg M.Z., Zhu S., Bahri N., Kyriazoglou A., Xu L., Chen T., Mariño-Enriquez A, & Fletcher J.A. (2021). YWHAE-NUTM2 oncoprotein regulates proliferation and cyclin D1 via RAF/MAPK and Hippo pathways. Oncogenesis, 10(5), 37.

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Optimized CD4i Fusion Protein Production

Cited 1 time

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All CD4i fusion constructs were codon optimized and cloned into pcDNA3.1 (–) using EcoRI and HindIII (GenScript, Piscataway, NJ). The proteins were expressed by transient transfection in 293F cells. Supernatants were harvested 5 days post transfection, filtered, and purified by cOmplete His-tag purification resin (Roche, Basel, Switzerland). The RSC3 core (11 (link)), BG505.SOSIP.664 trimer (48 (link)) with their CD4bs knock out variants, and 16055 NFL TD CC ΔGly4 bearing 4 glycan deletions (residues 276, 301, 360, & 463) (49 (link)) were purified as described previously.
To remove aggregates and undesired oligomeric states, all protein samples were subject to size exclusive chromatography (SEC) on Superpose6 10/300GL or Superdex 200 columns (GE Healthcare) pre-equilibrated with PBS in an ÄKTA Pure Station. Protein samples purity was analyzed by SDS PAGE.

Galkin A., Chen Y., Guenaga J., O’Dell S., Acevedo R., Steinhardt J.J., Wang Y., Wilson R., Chiang C.I., Doria-Rose N., Grishaev A.V., Mascola J.R, & Li Y. (2020). HIV-1 gp120-CD4i Antibody Complex Elicits CD4 Binding Site Specific Antibody Response in Mice. Journal of immunology (Baltimore, Md. : 1950), 204(6), 1543-1561.

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