ArticleGenome research2004
High-throughput generation of P. falciparum functional molecules by recombinational cloning.
Article in Genome research, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed, 62 citations in OpenAlex.
- Discovery of Novel Plasmodium falciparum Pre-Erythrocytic Antigens for Vaccine Development.PloS one · 2015Trial
- Antibody responses in Burkinabe children againstFrontiers in immunology · 2025Article
- Perkinsus marinus in bioreactor: growth and a cost-reduced growth medium.Journal of industrial microbiology & biotechnology · 2023Article
- Generating genomic platforms to study Candida albicans pathogenesis.Nucleic acids research · 2018Article
- Identification of target proteins of clinical immunity to Plasmodium falciparum in a region of low malaria transmission.Parasitology international · 2018Article
- Review
- Review
- Progress with viral vectored malaria vaccines: A multi-stage approach involving "unnatural immunity".Vaccine · 2015Review
- Malaria invasion ligand RH5 and its prime candidacy in blood-stage malaria vaccine design.Human vaccines & immunotherapeutics · 2015Review
- Exploration of panviral proteome: high-throughput cloning and functional implications in virus-host interactions.Theranostics · 2014Article
- A library of functional recombinant cell-surface and secreted P. falciparum merozoite proteins.Molecular & cellular proteomics : MCP · 2013Article
- Review
- Measurement of naturally acquired humoral immune responses against the C-terminal region of the Plasmodium vivax MSP1 protein using protein arrays.Parasitology research · 2011Article
- Production of Myxoma virus gateway entry and expression libraries and validation of viral protein expression.Current protocols in microbiology · 2011Article
- Identification of two new protective pre-erythrocytic malaria vaccine antigen candidates.Malaria journal · 2011Article
- Plasmodium immunomics.International journal for parasitology · 2011Review
- Evaluation of approaches to identify the targets of cellular immunity on a proteome-wide scale.PloS one · 2011Article
- Insect cells are superior to Escherichia coli in producing malaria proteins inducing IgG targeting PfEMP1 on infected erythrocytes.Malaria journal · 2010Article
- Production of recombinant proteins from protozoan parasites.Trends in parasitology · 2010Review
- Genetic manipulation of francisella tularensis.Frontiers in microbiology · 2010Article
Corrections and comments
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Authors and funding
15 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Large-scale functional genomics studies for malaria vaccine and drug development will depend on the generation of molecular tools to study protein expression. We examined the feasibility of a high-throughput cloning approach using the Gateway system to create a large set of expression clones encoding Plasmodium falciparum single-exon genes. Master clones and their ORFs were transferred en masse to multiple expression vectors. Target genes (n = 303) were selected using specific sets of criteria, including stage expression and secondary structure. Upon screening four colonies per capture reaction, we achieved 84% cloning efficiency. The genes were subcloned in parallel into three expression vectors: a DNA vaccine vector and two protein expression vectors. These transfers yielded a 100% success rate without any observed recombination based on single colony screening. The functional expression of 95 genes was evaluated in mice with DNA vaccine constructs to generate antibody against various stages of the parasite. From these, 19 induced antibody titers against the erythrocytic stages and three against sporozoite stages. We have overcome the potential limitation of producing large P. falciparum clone sets in multiple expression vectors. This approach represents a powerful technique for the production of molecular reagents for genome-wide functional analysis of the P. falciparum genome and will provide for a resource for the malaria resource community distributed through public repositories.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.