Team:Tunghai TAPG/Parts
Antimicrobial peptide has been in the world for a while, but still couldn’t be common for people; there are several reasons, for instance, high prices, the production time is too long, the techniques couldn’t be accepted by people, etc. While there are various peptide companies, but most of peptides are synthesized by the chemical method, it takes lots of time, and the costs is high, e.g. 1 mg of peptide may cost approximately US $25-30. To overcome the difficulties of the peptide company, we are using the techniques of genetic engineering to synthesize antimicrobial peptide. In this combination of biobrick, our final goal is to use an efficient, convenience way to produce our antimicrobial peptide. In order to achieve the goal, we have constructed a recombinant plasmid that could express a fusion protein containing the fragment of our antimicrobial peptide. For example, his tag that could make purification more easy, sumo that could increase the solubility of fusion protein, and the t7 promoter that has the higher specificity, etc.
Technically, the current manufacture of peptide could be split into two categories: chemical method and genetic engineering method. For the peptide that has a shorter sequence or the small-scale production, chemical method has the advantages, because of the foundation that built from synthetic chemical factors. However, as the peptide we need to synthesize is longer, the accuracy from the chemical approach would be decreased. Although, the construction of an applicable recombinants DNA is difficult at the initial stage, which requires addition costs and efforts, once the expression and purification processes are successful, the production can be easily scaled up, with a much lower overall costs; Factories could use this technique to establish an industrial procession, including bacteria cultivation, expression, disruption, purify, these all could be systematic in factories, thus, using the technique of recombinant DNA for peptide production could be a great expectation of peptides and peptide drugs development.
BBa_k2920001
| Name of the Part | Type | Description | Length (bp) |
|---|---|---|---|
| BBa-K2920727 | Protein_Domain | We can prove our gene has the antimicrobial functions after we transform the constructed plasmid containing the peptide gene, so we can achieve the mass production of our specific peptide by genetic engineering approach and develop further applications. | 33bp |
| BBa- K1911005 | Coding | eGFP =eGFP = Enhanced green fluorescent protein which can be used as a reporter protein. eGFP was used in our project to show the effects of attaching the degradation tag to a target gene. In essence, the eGFP should be less prominent in E. coli that contain the tags compared to the E. coli cells that have been constructed, which do not have any degradation tags attached to the eGFP. eGFP can be used as a reporter protein and helps the observer easily see the coloration of E. coli cells which indicate a successful transformation. | 717bp |
| BBa- I719005 | Regulatory | (T7 Promoter) (T7 Promoter) The constitutive promoter derived from the T7 bacteriophage allows a high expression of proteins only when the T7 polymerase is present. | 23bp |
| BBa- J18909 | tag | (His 6x tag) This codes for six Histidine residues to allow for His column protein purification. It will work best if put at either the N or C terminus of your protein construct. | 18bp |
| BBa- K2323012 | tag | (SUMO tag) The SUMO tag increases the solubility of the tagged protein and can be cleaved with the SUMO protease. | 297bp |
| BBa- K731721 | terminator | (T7 terminator)a wild type terminator from T7 bacteriophage | 48bp |
| BBa- K1638034 | coding | (Sce VMA Intein)This part consists of an intein from Saccharomyces cerevisiae VMA1 gene including a CBD-tag. This intein is a thiol-induced self-cleavable protein that enables the release of a C-terminal fused protein. The chitin-binding domain is an affinity tag that enables affinity purification on a chitin column. First, the target protein fused to intein is loaded and washed on the chitin column. Using a thiol reagent, like dithiothreitol (DTT), an on-column cleavage is induced and the target protein is released. | 1534bp |