Part:BBa_K4006003
Ferritin
This part includes a coding sequence for the protein ferritin codon optimized for use in the Chlamydomonas reinhardtii chloroplast. Ferritin is a large protein that takes the form of a hollow sphere, sequestering iron inside of itself. While it is extremely well known as a tool for iron sequestration, it can be used to capture multiple heavy metals. It has also been expressed within the Chlamydomonas reinhardtii chloroplast before with a notable difference in capacity for binding iron. We were able to successfully clone the part into our plasmid and select transformed colonies in E. coli, but we did not obtain any transformant C. reinhardtii colonies after biolistic transformation.
We were able to clone this construct into our plasmid, pASapI, using Gibson assembly and select for transformed E. coli colonies.
PLATE PICTURES
Digestion of the miniprepped DNA in the plasmid pASapI with XbaI and BstXI should result in two bands of approximate sizes CHANGE SIZE bp and CHANGE SIZE bp as compared to the original plasmid which should have three bands of sizes 4446, 1376, and 800 bp. Each of the colonies were successfully cloned.
GEL PICTURE
Unfortunately, we were unable to successfully transform this construct into C. reinhardtii and test expression or effect on arsenic uptake in the algae.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 5
Illegal SapI.rc site found at 515
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