Part:BBa_K3264005
NT4RepCT
Spider silk serves as a new material with superior properties that can be applied in medication, cloth, and aerospace fields. However, spider breeding is not applicable due to spider's fierce behavior. The current approach is to produce recombinant spidroins (silk proteins) from other chassis and spin them into silk. NT-2Rep-CT is the common architecture in two main types of spider silk proteins: major ambulate spidorins(MaSps) and minor ampullate spidorins (MiSps). NT is a non-repetitive N-terminal domain, as well as CT, the C-terminal domain. In between them is an extensive repetitive region-Rep In the storage of spider silk protein, NT is a monomeric and highly soluble region in neutral pH, and it provides the solubility of the entire spidroin. NT can form stable dimers when pH decreases in the spinning duct. On the other side during the decrease of pH, the CT gets disrupted and unfold to turn in the beta-sheet amyloid-like fibrils. It is hypothesized that structural change of CT precipitate the change of the repetitive region to beta-sheet conformation. Enable to get a step closer to unobtainable size (556 kDa), containing 192 repeat motifs of the Nephila clavipes dragline spidroin, we have developed a synthetic biology approach where we combine standardized DNA part assembly and split the intermediated ligation to produce recombinant spidroins of NT-4Rep-CT, in this case, extra a 2Rep is added to the NT-2Rep-CT and was expected to have a better mechanical performance after spinning.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 943
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 424
Illegal SpeI site found at 943 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 943
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 943
- 1000COMPATIBLE WITH RFC[1000]
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