Our goal is to be able to implement Iterative Capped Assmebly (ICA) for use in designing custom spider silk genes. The spider silk protein has a highly repetitive primary structure, and its properties can be changed by changing the sequence and identities of these repeats. We want to be able to fully control the primary sequence of our silk gene, and develop a quick, efficient method of customizing assembly of the spider-silk genes.
We successfully constructed biobricks that contain our silk genes and the sticky overhangs. 4/24/2015
We proved that our designed sticky ends work as expected, and exhibit specific binding, showing that ICA can work in spider silk genes. 5/8/2015 and 5/12/2015
We have established a protocol generating fragments to be used in ICA.
We successfully characterized the use of ICA oligos with MaSp AB, BC, CA and showed that they could work in ICA! 6/3/2015
We successfully made a 3-mer using the ICA protocol on M-270 Streptavidin Beads! 7/7/2015
We are working on optimizing PCR amplification of MaSp and BsaI digestion so as to get the most amount of working monomers for ICA.--DONE! see here
We need to characterize our MaSp monomers and how they behave when used with the initiators, terminators, and capping oligos that are also used in ICA.--DONE! 6/3/2015
We are going to start actual ICA soon.--STARTED! see 6/19/2015
We are optimizing ICA to get it to work just right!
Our final goal is to make a several different 15-mers that have different rations of MaSp1 and MaSp2 that we can express and characterize.
