Part:BBa_K1919001
Constitutive promotor J23105 and downstream CecropinXJ
Biobrick BBa_K1919001 is a composite part, consisting of constitutive promotor J23105, RBS, TrxA tag, 6xHis tag, thrombin site, S tag, enterkinase site and the coding sequence of antimicrobial peptide CecropinXJ.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 506
Illegal BamHI site found at 549
Illegal XhoI site found at 750 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Biology
Antimicrobial peptides (AMPs) are a group of peptides that play roles in the innate immune system to protect the host from invading pathogens [1]. AMPs have minimal toxicity and low sensitivity effects to the host [2], which means antimicrobial peptides have the potential to be used to replace antibiotics in the future. Thus, the detrimental effects of antibiotics overuse will be released.
Cecropins, a group of small AMPs mainly found in the hemolymph of insects, consist of 31 39 amino acid residues and have a broad spectrum, high heat stability and potent bacteriostatic activity [3-5]. CecropinXJ (Part BBa_K1919000) is a member of the Cecropin family, which was first cloned from the larvae of the Xinjiang silkworm (Bombyx mori). Previous researches have determined the complete amino acid sequence of this molecule [6]. It has been demonstrated that CecropinXJ could be expressed in eukaryotic expression system such as Pichia pastoris [7] or prokaryotic expression system such as E.coli [8]. What’s more, CecropinXJ exhibited to have various activities such as antibacterial activity against both Gram‑positive and Gram-negative bacteria, as well as antifungal activity [8]. These characteristics indicate that CecropinXJ is an ideal antimicrobial substance to be used to treat foot diseases caused by microbes.
Results
This part is constructed based on part BBa_K1919000 [1], which express cecropinXJ continuously downstream of constitutive promoter J23105 [2](Fig. 1)
The combination of cecropinXJ and constitutive promoters were determined by PCR. (Fig. 2) However, there is no clear band being detected while using SDS-PAGE and Western blot, which might due to the high level expression of AMP inhibited the growth of host itself.
//function/odor
biology | Bombyx mori |
chassis | E.coli |
protein | TrxA+6xHis tag+Thrombin site+S Tag+enterokinase site+CecropinXJ |