Difference between revisions of "Part:BBa K3113101"
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mGag is short for mouse Gag. This sequence codes for the coat protein of the human immunodeficiency virus optimised to bud in mice. It mediates the essential events in virion assembly, including binding the plasma membrane, making the protein-protein interactions necessary to create spherical particles. | mGag is short for mouse Gag. This sequence codes for the coat protein of the human immunodeficiency virus optimised to bud in mice. It mediates the essential events in virion assembly, including binding the plasma membrane, making the protein-protein interactions necessary to create spherical particles. | ||
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| + | While exosome secretion is a ubiquitous mechanism, HIV Gag vesicle formation is a bioorthogonal mechanism in mice. This poses a problem because it is known that small animal models have several cellular barriers to prevent HIV replication. In mice, assembly and budding of HIV vesicles are blocked<ref>Sherer, N.M., Swanson, C.M., Papaioannou, S., and Malim, M.H. (2009). Matrix Mediates the Functional Link between Human Immunodeficiency Virus Type 1 RNA Nuclear Export Elements and the Assembly Competency of Gag in Murine Cells. J. Virol. 83, 8525–8535. | ||
| + | </ref>, thus severely interfering with the usage of Gag vesicles for our purposes. However, publications report different modifications to overcome these limitations<ref>Quellen</ref>. We, therefore, codon-optimized our Gag construct for expression in mouse cells and introduced the point mutation L21S<ref>Diaz-Griffero, F., Taube, R., Muehlbauer, S.M., and Brojatsch, J. (2008). Efficient production of HIV-1 viral-like particles in mouse cells. Biochem. Biophys. Res. Commun. 368, 463–469.</ref> to aid vesicle formation and secretion. | ||
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<partinfo>BBa_K3113101 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3113101 SequenceAndFeatures</partinfo> | ||
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Latest revision as of 09:23, 12 December 2019
mGag
mGag is short for mouse Gag. This sequence codes for the coat protein of the human immunodeficiency virus optimised to bud in mice. It mediates the essential events in virion assembly, including binding the plasma membrane, making the protein-protein interactions necessary to create spherical particles.
Usage and Biology
While exosome secretion is a ubiquitous mechanism, HIV Gag vesicle formation is a bioorthogonal mechanism in mice. This poses a problem because it is known that small animal models have several cellular barriers to prevent HIV replication. In mice, assembly and budding of HIV vesicles are blocked[1], thus severely interfering with the usage of Gag vesicles for our purposes. However, publications report different modifications to overcome these limitations[2]. We, therefore, codon-optimized our Gag construct for expression in mouse cells and introduced the point mutation L21S[3] to aid vesicle formation and secretion.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 779
Illegal BglII site found at 1307 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 709
- 1000COMPATIBLE WITH RFC[1000]
References
- ↑ Sherer, N.M., Swanson, C.M., Papaioannou, S., and Malim, M.H. (2009). Matrix Mediates the Functional Link between Human Immunodeficiency Virus Type 1 RNA Nuclear Export Elements and the Assembly Competency of Gag in Murine Cells. J. Virol. 83, 8525–8535.
- ↑ Quellen
- ↑ Diaz-Griffero, F., Taube, R., Muehlbauer, S.M., and Brojatsch, J. (2008). Efficient production of HIV-1 viral-like particles in mouse cells. Biochem. Biophys. Res. Commun. 368, 463–469.