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Induction of Protein Expression Within Escherichia coli Vector for Entry into Mammalian Cells
- Source :
- Human Gene Therapy Methods. 25:40-47
- Publication Year :
- 2014
- Publisher :
- Mary Ann Liebert Inc, 2014.
-
Abstract
- Direct protein delivery into the cytosol of mammalian cells by invasive Escherichia coli (E. coli) bacterial vector will bypass the need to achieve nuclear entry and transcription of DNA, a major hurdle that is known to seriously limit gene transfer. The bacterial vector is induced to express the protein during its growth phase, before presentation for entry into mammalian cells and release of its content into the cellular environment. For this class of vector, crossing the plasma membrane becomes the primary step that determines the success of protein delivery. Yet, how the mechanics of protein expression within the vector affect its entry into the host is poorly understood. We found the vector's effectiveness to enter HeLa cells diminished together with its viability when phage N15 protelomerase (TelN) expression was induced continuously in the invasive E. coli despite producing an abundant amount of functional protein. By comparison, shorter induction, even as little as 3 hr, produced sufficient amounts of functional TelN and showed more effective invasion of HeLa cells, comparable to that of uninduced invasive E. coli. These results demonstrate that brief induction of protein expression during vector growth is essential for optimal entry into mammalian cells, an important step for achieving bacteria-mediated protein delivery.
- Subjects :
- Time Factors
Growth phase
Gene Expression
Gene transfer
Biology
medicine.disease_cause
Applied Microbiology and Biotechnology
Protein expression
HeLa
Viral Proteins
chemistry.chemical_compound
Transcription (biology)
Escherichia coli
Genetics
medicine
Humans
Telomerase
Genetics (clinical)
Pharmacology
Enzyme Precursors
Gene Transfer Techniques
Tetracycline
biology.organism_classification
Molecular biology
Cell biology
Cytosol
chemistry
Molecular Medicine
DNA
HeLa Cells
Plasmids
Subjects
Details
- ISSN :
- 19466544 and 19466536
- Volume :
- 25
- Database :
- OpenAIRE
- Journal :
- Human Gene Therapy Methods
- Accession number :
- edsair.doi.dedup.....ed18517f3b939781c4c015c74043f7d6