Difference between revisions of "Part:BBa K3733007"

Revision as of 13:50, 20 October 2021

LTA: A novel antimicrobial peptide

LTA is a novel antimicrobial peptide (AMP), whose design was based on combing the active centers of a ride range of AMPs, including LL-37, YW12D, innate defense regulator 1, and cathelicidin 2 with thymopentin or the active center of thymosin alpha 1 (Tα1). It could neutralize Lipopolysaccharides (LPS), thus effectively blocking the downstream inflammation pathway.

Design

This part has undergone codon optimization based on the bias of Escherichia coli. As the original coding sequence of LTA doesn’t contain the start codon and termination codon, they are then added to guarantee the expression of the protein.

Materials and Method

1.Plasmids Construction: LTA-coding sequence is constructed using primers by overlap extension PCR. The constructed fragment is ligated to the target site in pET28 by PCR and homologous recombination. The correct construction is confirmed by sequencing.

2.Expression and Purification

    1.Plasmid pET-28a(+)-LTA (with His-tag) is transformed to Escherichia coli BL21(DE3). The E. coli strain is cultured in LB medium containing 10 μg/mL kanamycin.

Sequence and Features