Difference between revisions of "Part:BBa K3739041"

Revision as of 21:47, 21 October 2021

J23100-B0030-LMT-his-GFP-B0010

This part contains a Vibrio natriegens-optimized GFP coding sequence, and the GFP is fused at N-terminal with his-tag in order to let us purify the protein. His-GFP is fused with secretory peptide from LMT.We use BBa_K3739041 to construct the expression system and help prove the function of some signal peptide.

Biology

LMT
Lytic murein transglycosylase (LMT) is an enzyme which is able to degrade murein, a component of cell wall of bacteria. There are two kind of LMTs existing in E.coli: the membrane-binding one and the soluble one. The gene coding LMT homolog is also incorporated in the genome of Vibrio natriegens. The LMT signal peptide (named LMT in our parts) is from the LMT homolog, which can lead the fused protein secreted out of Vibrio natriegens.
GFP
Protein tags are peptides genetically fused to the proteins of interest. There are many different kinds of tags developed for for different purposes. For labeling experiments, normally the proteins of interest are tagged directly by fluorescent tags. Green fluorescent protein (GFP) make it easier to study protein localization, interactions and dynamics when fusing with other peptides and proteins.

Usage

Here, we used BBa_K3739010 to construct the expression system and obtained the composite part BBa_K3739041. We transformed the constructed plasmid on the expression vector pET-28a (+) into V. natriegens to extract the protein. This protein work together with those from BBa_K3739038 and BBa_K3739043 to verify the function of our secretory peptides Aly01 and LMT.

Characterization

Sequence and Features