Part:BBa_K4339001
Alanine tRNA synthetase
Alanine tRNA synthetase is an enzyme that catalyses the binding of alanine to tRNA. Modelling demonstrated that the expression of spider silk proteins (spidroins), which have a number of repetitive alanine regions, would be increased with the co-expression of alanine tRNA synthetase.
Usage and Biology
Sequence and Features
The sequence originates from E. coli MG1655 K-12 (Puteny et al. 1981)
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 2066
Illegal PstI site found at 2582 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 2066
Illegal PstI site found at 2582 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 629
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 2066
Illegal PstI site found at 2582 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 2066
Illegal PstI site found at 2582
Illegal AgeI site found at 709 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1090
Illegal SapI.rc site found at 1138
Illegal SapI.rc site found at 1288
The enzyme was intended for co-expression with MaSps (Major Ampullate Spidroin Proteins) as part of a helper plasmid, containing genes for alanine tRNA and CycA (part K4339000 - an alanine import channel protein) expression. This system was designed to increase alanine uptake and synthesis of alanyl-tRNA to maximise possible rate of alanine incorporation into protein and thus, production of alanine-rich MaSps.
This part was successfully synthesised but was not transformed due to time constraints.
References
Putney S et al. Purification and properties of alanine tRNA synthetase from Escherichia coli. A tetramer of identical subunits. Journal of Biological Chemistry. 1981;256(1): 198–204. doi: https://doi.org/10.1016/s0021-9258(19)70119-7
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