Difference between revisions of "Part:BBa K4759009:Experience"
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===Applications of BBa_K4759009=== | ===Applications of BBa_K4759009=== | ||
| + | Superfolder GFP is a basic (constitutively fluorescent) green fluorescent protein published in 2005, derived from Aequorea victoria. It is reported to be a very rapidly-maturing weak dimer | ||
| + | Molecular engineering of the green fluorescent protein (GFP) into a robust and stable variant named Superfolder GFP (sfGFP) has revolutionized the field of biosensor development and the use of fluorescent markers in diverse area of biology. sfGFP-based self-associating bipartite split-FP systems have been widely exploited to monitor soluble expression in vitro, localization, and trafficking of proteins in cellulo. In our research, we use split this part into three parts——GFP1-9、GFP10、GFP11，we use linker to link GFP1-9、GFP10 to ferredoxin，link GFP11 to OleP. | ||
| + | The 1st – 10th β fold (simply gfp1-10) separates the 11th β fold in the sf gf p structure from the 11th (gfp11), each individually unable to fluoresce, and when gfp1-10 is mixed with a protein containing gfp11 fragment, gfp1-10 actively binds to gfp11 (indicating the higher affinity of gfp1-10 and gfp11) to restore fluorescence activity | ||
| + | The main principle of bifc, is the fluorescent protein molecules into two separate cannot fluorescent part, respectively fusion with the target protein, when two target protein interact close to each other, the two fluorescent molecule fragments because of physical distance, and complementary, the formation of active fluorescent protein can be detected. Therefore, this fluorescence signal due to complementarity and its strength can be used to indicate whether the two target proteins interact and their strength | ||
| + | When gfp10 and gfp11 were fused to the target proteins a and b, respectively, and then the two fusion proteins were coexpressed in gfp1-9, due to the affinity between gfp1-9 and gfp11, when gfp10 and gfp11 interact through the target protein, gfp1-9 was also recruited nearby because of the affinity with gfp11, and the three segments were repooled to restore fluorescence activity. This tripartite sf gf p system can be used for protein interaction detection. In addition to the intuitive advantages, the gfp10 and gfp11 are both short peptides, so the influence on the fusion protein structure is probably small. In addition, because it is divided into three parts, the fluorescence background is further reduced, which helps to improve the signal-to-noise ratio of the detection. | ||
===User Reviews=== | ===User Reviews=== | ||
Revision as of 15:05, 7 October 2023
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Applications of BBa_K4759009
Superfolder GFP is a basic (constitutively fluorescent) green fluorescent protein published in 2005, derived from Aequorea victoria. It is reported to be a very rapidly-maturing weak dimer Molecular engineering of the green fluorescent protein (GFP) into a robust and stable variant named Superfolder GFP (sfGFP) has revolutionized the field of biosensor development and the use of fluorescent markers in diverse area of biology. sfGFP-based self-associating bipartite split-FP systems have been widely exploited to monitor soluble expression in vitro, localization, and trafficking of proteins in cellulo. In our research, we use split this part into three parts——GFP1-9、GFP10、GFP11，we use linker to link GFP1-9、GFP10 to ferredoxin，link GFP11 to OleP. The 1st – 10th β fold (simply gfp1-10) separates the 11th β fold in the sf gf p structure from the 11th (gfp11), each individually unable to fluoresce, and when gfp1-10 is mixed with a protein containing gfp11 fragment, gfp1-10 actively binds to gfp11 (indicating the higher affinity of gfp1-10 and gfp11) to restore fluorescence activity The main principle of bifc, is the fluorescent protein molecules into two separate cannot fluorescent part, respectively fusion with the target protein, when two target protein interact close to each other, the two fluorescent molecule fragments because of physical distance, and complementary, the formation of active fluorescent protein can be detected. Therefore, this fluorescence signal due to complementarity and its strength can be used to indicate whether the two target proteins interact and their strength When gfp10 and gfp11 were fused to the target proteins a and b, respectively, and then the two fusion proteins were coexpressed in gfp1-9, due to the affinity between gfp1-9 and gfp11, when gfp10 and gfp11 interact through the target protein, gfp1-9 was also recruited nearby because of the affinity with gfp11, and the three segments were repooled to restore fluorescence activity. This tripartite sf gf p system can be used for protein interaction detection. In addition to the intuitive advantages, the gfp10 and gfp11 are both short peptides, so the influence on the fusion protein structure is probably small. In addition, because it is divided into three parts, the fluorescence background is further reduced, which helps to improve the signal-to-noise ratio of the detection.
User Reviews
UNIQ093a9dc1123d0b71-partinfo-00000000-QINU UNIQ093a9dc1123d0b71-partinfo-00000001-QINU