Part:BBa_K1621003
TeNT_Hc - C-terminal part of the heavy chain (tetanus toxin)
This part contains a coding sequence belonging to the tetanus neurotoxin. It is not responsible for the toxic properties and therefore exhibits no toxicity in humans.
The source organism of this part is Clostridium tetani, an anaerobic, gram-positive bacterium causing tetanus disease. C. tetani produces two different toxins, tetanolysin and tetanospasmin. The first is responsible for damage of the heart muscle and blood components. The latter causes the more prominent symptoms like hypersensitivity, increased reactions and spasms.
The neurotoxin (or tetanospasmin) is composed of a heavy and a light chain. They are translated as one polypeptide of about 150 kDa and cleaved by a protease afterwards. The resulting heavy (~100 kDa) and light chain (~50kDa) are linked to each other by two disulfide bonds (Yu et al., 2011).
Regarding the functions, the light chain is the toxic part of the toxin that is translocated into the neuron at the neuromuscular junction. The heavy chain’s function is to mediate this translocation. Therefore, the C-terminal part interacts with the cells membrane via specific receptors and the N-terminal part aids in the translocation process itself. In the cell, the light chain prevents the exocytosis of neurotransmitters (Rummel et al., 2003).
Nonetheless, the C-terminal part of the heavy chain which is encoded by this part is promising in the development of second generation vaccines against the toxin itself (Yu et al., 2011). This means of course, that it provokes an immune response in humans and work has to be done under consideration of the respective safety requirements.
As Yu et al. (2011) showed before, cloning the sequence into an expression vector enables efficient overexpression of the part. [Results will be updated]
The part has been shipped to the registry in standard pSB1C3 and begins with a start codon (ATG). Cloning into the chipping backbone was performed by Gibson Assembly and the sequence was verified afterwards.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 430
- 1000COMPATIBLE WITH RFC[1000]
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