ArticleBMC biotechnology2008
Construction of a series of vectors for high throughput cloning and expression screening of membrane proteins from Mycobacterium tuberculosis.
Article in BMC biotechnology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers.
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Who cites it
33 citing papers in PubMed, 47 citations in OpenAlex.
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- Functional comparison of full-length palladin to isolated actin binding domain.Protein science : a publication of the Protein Society · 2023Article
- Functional Tolerance to Cysteine Mutations in Human α7 Nicotinic Acetylcholine Receptors.ACS chemical neuroscience · 2020Article
- Myopalladin promotes muscle growth through modulation of the serum response factor pathway.Journal of cachexia, sarcopenia and muscle · 2020Article
- Article
- Efficacy of mucosal polyanhydride nanovaccine against respiratory syncytial virus infection in the neonatal calf.Scientific reports · 2018Article
- Human oncoprotein Musashi-2 N-terminal RNA recognition motif backbone assignment and identification of RNA-binding pocket.Oncotarget · 2017Article
- Functional Human α7 Nicotinic Acetylcholine Receptor (nAChR) Generated from Escherichia coli.The Journal of biological chemistry · 2016Article
- Computational modeling of TC0583 as a putative component of the Chlamydia muridarum V-type ATP synthase complex and assessment of its protective capabilities as a vaccine antigen.Microbes and infection · 2016Article
- Actin polymerization is stimulated by actin cross-linking protein palladin.The Biochemical journal · 2016Article
- Production of Well-Characterized Virus-like Particles in an Escherichia coli-Based Expression Platform for Preclinical Vaccine Assessments.Methods in molecular biology (Clifton, N.J.) · 2016Article
- Hypothetical protein CT398 (CdsZ) interacts with σ(54) (RpoN)-holoenzyme and the type III secretion export apparatus in Chlamydia trachomatis.Protein science : a publication of the Protein Society · 2015Article
- Characterization of Dye-decolorizing Peroxidase (DyP) from Thermomonospora curvata Reveals Unique Catalytic Properties of A-type DyPs.The Journal of biological chemistry · 2015Article
- KU675, a Concomitant Heat-Shock Protein Inhibitor of Hsp90 and Hsc70 that Manifests Isoform Selectivity for Hsp90α in Prostate Cancer Cells.Molecular pharmacology · 2015Article
- Membrane protein structure from rotational diffusion.Biochimica et biophysica acta · 2015Review
- Actin-induced dimerization of palladin promotes actin-bundling.Protein science : a publication of the Protein Society · 2015Article
- Recombinant expressed vector pET32a (+) S constructed by ligation independent cloning.Molecules (Basel, Switzerland) · 2014Article
- Assignment of oriented sample NMR resonances from a three transmembrane helix protein.Journal of magnetic resonance (San Diego, Calif. : 1997) · 2014Article
- Chlamydia trachomatis CT771 (nudH) is an asymmetric Ap4A hydrolase.Biochemistry · 2014Article
- Lipid bilayer preparations of membrane proteins for oriented and magic-angle spinning solid-state NMR samples.Nature protocols · 2013Article
Corrections and comments
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Authors and funding
6 authors at 3 institutions in 1 country.
Funding
Abstract
backgroundOne of the major challenges for membrane protein structural genomics is establishing high-throughput cloning and expression screening methods to obtain enough purified protein in a homogeneous preparation for structural and functional studies. Here a series of ligation independent cloning based vectors were constructed to address this challenge.
resultsThe feasibility of these vectors was tested with 41 putative membrane proteins from Mycobacterium tuberculosis. The efficiency for direct cloning of these target genes from PCR products was 95% (39/41). Over 40% of cloned genes were overexpressed in Escherichia coli BL21 (DE3)-RP codon plus strain in the first round of expression screening. For those proteins which showed no expression, three protein fusion partners were prepared and it was found that each of the target proteins could be overexpressed by at least one of these fusions, resulting in the overexpression of two thirds of the cloned genes.
conclusionThis expression platform features high throughput cloning, high flexibility for different constructs, and high efficiency for membrane protein overexpression, and is expected to be useful in membrane protein structural and functional studies.
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