Protein synthesis in liposomes with a minimal set of enzymes
In a significant step towards the construction of the semi-synthetic minimal cell, a protein expression system with a minimal set of pure and specific enzymes is required. A novel cell-free transcription and translation system named PURESYSTEM (PS), consisting of a specified set of 36 enzymes and ri...
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| Veröffentlicht in: | Biochemical and biophysical research communications 2007-11, Vol.363 (1), p.12-17 |
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| container_title | Biochemical and biophysical research communications |
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| creator | Murtas, Giovanni Kuruma, Yutetsu Bianchini, Paolo Diaspro, Alberto Luisi, Pier Luigi |
| description | In a significant step towards the construction of the semi-synthetic minimal cell, a protein expression system with a minimal set of pure and specific enzymes is required. A novel cell-free transcription and translation system named PURESYSTEM (PS), consisting of a specified set of 36 enzymes and ribosomes, has been entrapped in POPC liposomes for protein synthesis. The PS has been used to transcribe and translate an Enhanced Green Fluorescent Protein (EGFP) gene from plasmid DNA. The synthesis is confirmed by the EGFP fluorescence emitting liposomes on fluorometric analysis and on confocal microscopy analysis. Furthermore the PS encapsulated into POPC liposomes can drive the expression of the
plsB and
plsC genes encoding for the
sn-glycerol-3-phosphate acyltransferase (GPAT) and 1-acyl-
sn-glycerol-3-phosphate acyltransferase (LPAAT) involved in the first step of the “salvage pathway” for synthesis of POPC. The expression of GPAT and LPAAT in liposomes would in principle allow the production of the cell boundary from within. |
| doi_str_mv | 10.1016/j.bbrc.2007.07.201 |
| format | Article |
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plsB and
plsC genes encoding for the
sn-glycerol-3-phosphate acyltransferase (GPAT) and 1-acyl-
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plsB and
plsC genes encoding for the
sn-glycerol-3-phosphate acyltransferase (GPAT) and 1-acyl-
sn-glycerol-3-phosphate acyltransferase (LPAAT) involved in the first step of the “salvage pathway” for synthesis of POPC. 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A novel cell-free transcription and translation system named PURESYSTEM (PS), consisting of a specified set of 36 enzymes and ribosomes, has been entrapped in POPC liposomes for protein synthesis. The PS has been used to transcribe and translate an Enhanced Green Fluorescent Protein (EGFP) gene from plasmid DNA. The synthesis is confirmed by the EGFP fluorescence emitting liposomes on fluorometric analysis and on confocal microscopy analysis. Furthermore the PS encapsulated into POPC liposomes can drive the expression of the
plsB and
plsC genes encoding for the
sn-glycerol-3-phosphate acyltransferase (GPAT) and 1-acyl-
sn-glycerol-3-phosphate acyltransferase (LPAAT) involved in the first step of the “salvage pathway” for synthesis of POPC. The expression of GPAT and LPAAT in liposomes would in principle allow the production of the cell boundary from within.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>17850764</pmid><doi>10.1016/j.bbrc.2007.07.201</doi><tpages>6</tpages></addata></record> |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Biomimetics - methods Cell-Free System Cell-free transcription and translation Enzyme Activation Escherichia coli - enzymology Escherichia coli Proteins - chemistry Liposomes Liposomes - chemistry Minimal cell Minimal genome Multienzyme Complexes - chemistry Origin of life Protein synthesis Proteins - chemical synthesis Synthetic biology |
| title | Protein synthesis in liposomes with a minimal set of enzymes |
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