Part:BBa_K2455002:Design
CPP-SYFP2
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The first step in the insertion was amplification of SYFP2 using primers matching the USER Cassette of the Cell-Penetrating USER Cassette. To do this we used the following primers:
Forward primer: CGTGCGAUCA-ATGGTTAGCAAGGGCGAAG
Reverse primer: CACGCGAUGA-TTTATACAGCTCATCCATACCCAGGG
Subsequently the PCR product could be gel-extracted and ligated into the Cell-Penetrating USER Cassette biobrick.
Finally for the creation of a biobrick version the following set of primers were used:
Forward primer: ATCCGGAATTCGCGGCCGCTTCTAG-ATGCGTCGCCGTCGCCGTCGCCGTCGCCGTGAATGCGTGCGATC
Reverse primer: CCAATGCATTGGTTCTGCAGCGGCCGCTACTAGTATTATTA-ATGGTGATGGTGATGATGTAAG
Source
The biobrick is an improved version of the Yellow Fluorescent Protein 2 (SYFP2).
References
Chang, M. et al. (2005). Cellular Internalization of Fluorescent Proteins via Arginine-rich Intracellular Delivery Peptide in Plant Cells. PCP Plant and Cell Physiology, 46(3), pp. 482-8.
Eudes & Chugh (2008). Cell-penetrating peptides. Plant Signal Behav., 3(8), pp. 549–550.