BBa_K942012 1 BBa_K942012 proU+RiAFP+Term 2012-09-25T11:00:00Z 2015-05-08T01:13:48Z The Raghium inquisitor Anti Freeze Protein (RiAFP) is a 137 AA protein that prevents the growth of ice crystals. The proU promoter is from E.coli and works activating transcription when the bacterium is osmotically stressed. (Kristiansen E, 2011) Kristiansen, E. et all (2011) Structural characteristics of a novel antifreeze protein from the longhorn beetle Rhagium inquisitor. Insect Biochem Mol Biol. 41(2):109-17. This protein has an approximate weight of 13.7kDa and is better visualized in a SDS-PAGE with Tris-Tricine gel buffer, because of its low molecular weight. It can be purified by affinity with its 6xHis tag. An induction with 300mM of NaCl presents good expression. (Gasteiger E., 2005) About Safety The RiAFP has no reported toxicity or hazardousness, but its important to remember to have the basic biosafety precautions using a biosecurity chamber when handling any transformed strains. The inductor also presents no hazard as it is just common salt. false false _1209_ 0 13053 9 It's complicated false The RiAFP is expressed when around 300mM NaCl added. This part was designed by adding a 6xHis tag for affinity purification to the anti freeze protein from Raghium inquisitor. This part was codon optimized for its expression in E.coli. false Luis Mario Leal Garza BBa_K942012_sequence 1 aatatcactacccgcagcagggaaataattcccgccaaatagctttttatcacgcaaataatttgtggtgatctacactgatactctgttgcattattcgcctgaaaccacaatattcaggcgttttttcgctatctttgacaaaaaatatcaactttctcgatttgctctcagcccttatatcacgggaaattccggcgatttgctcgcatcaatattcatgccacatttgccatcaggggttgcctcagattctcagtatgttagggtagaaaaaagtgactatttccattgggtaatatatcgacatagacaaataaaggaatctttctattgcatggcaattaaattagaaattaaaaatctttataaaatatttggcgagcatccacagcgagcgttcaaatatatcgaacaaggactttcaaaagaacaaattctggaaaaaactgggctatcgcttggcgtaaaagacgccagtctggccattgaagaaggcgagatatttgtcatcatgggattatccggctcgggtaaatccacaatggtacgccttctcaatcgcctgattgaacccacccgcgggcaagtgctgattgatggtgtggatattgccaaaatatccgacgccgaactccgtgaggtgcgcagaaaaaagaggaggtatgtactcgtgccgcgcagttggtgtcgacggtcgtgcagttacggacattcagggtacctgtcatgctaaggctacgggtgccggtgcgatggcctctggcaccagtgaaccgggttccacctcaacggcaaccgctacgggtcgtggtgcaaccgcacgttcgaccagcacgggccgtggtaccgcgaccacgaccgccaccggcacggcatccgctacctcaaacgcgattggccagggtaccgcaacgaccacggctacgggctcggcgggcggtcgcgccaccggtagcgcaaccacgagctctagtgcttctcagccgacccagacgcaaaccatcacgggcccgggttttcaaaccgccaaatcattcgcacgcaacaccgcaaccacgaccgttaccgcatcgcaccatcatcaccatcactaaaaaaaaaaaccccgcccctgacagggcggggtttttttt igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z