BBa_K1927000

BBa_K1927000 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1927000
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Marthe J??rgensen
Date created: 2016-09-22 11:00:00
Date modified: 2016-10-14 01:24:20

Beta-lacatamase enzyme called blaNDM-1



Types
DnaRegion

Roles
CDS

Coding

Sequences BBa_K1927000_sequence (Version 1)

Description

Usage and biology:

During the last decade antibiotic resistance has grown not only in incidents but also in

awareness among the public. Rapid detection of these incidents is essential in the battle

against sophisticated bacteria. Beta lactamases are enzymes that provides one of the many

mechanisms of antibiotic resistance. Its for this reason regulary used among researchers as

a selection marker. Bacteria that has been subjected to a procedure where foreign DNA is

introduced, an antibiotic resistance gene is often use to check if colonies succsesfully has

taken up the DNA and expressed it.

An example of this is ampicillin which is a highly used selection marker. The need for a

selection marker in genetical engineering has been shown extremly usefull. By transforming

a bacteria with a particular gene plus an antibiotic resistance gene it allows the researcher to

know exactly which colonies that has taken up the DNA. By plating them out on agar plates

with a given antibiotic, only the bacteria with the resistance gene can grow. Cells that did not

manage to take up the DNA will die.

The mechanism of beta lactamases are oriented to the bacterial cell wall. This cell wall is

unique to bacteria and consist of several components. Gram Positive and gram negative

bacteria will have a different cell wall composition.

In general, Gram??positive bacteria have a thicker layer of cell wall as well as a layer of

cytoplasmic membrane. These layers consist of several conserved compounds such as

monomeric disaccharide tetrapeptide, which are usually also those that will trigger an

immunological defence respons of the host.

Gram??negative bacteria (e.g., Escherichia coli) typically contain an outer membrane, an

intervening periplasmic space where a thin layer of cell wall resides, and a layer of

cytoplasmic membrane.

Beta lactamases are usually produced both by gram negative and positive bacteria, either

from plasmid or chromosomally. Beta lactamases are able to resist several types of

antibiotics. These antibiotics all have in common a 4 ?? atom ring called beta lactam ring

which the enzyme are able to hydrolyze and break open and the molecule looses its

antibacterial function.

Penicillin, a regulary used antibiotic have such a beta lactam ring.

This drug was the first antibiotic to be discovered and is still widely used today. This ring will

bind to an enzyme (DD ???transpeptidase) that is in charge of renewing the bacterial cell wall.

Without this enzyme there will be no new formations of peptidoglycans for the cell wall and

the integrity of the bacterial cell wall will be lost, it will eventually rupture and the bacteria will

die. By hydrolyzing the ring, it will make the molecule unable to bind to the cell wall

producing enzyme, thus the Penicillin have lost its destructive activity.

Notes

We added prefix and suffix to the gene so that we could perform restriction digestion and Gibson Assembly. We had to design the gene so that it will get the correct readingframe.

Source

The sequence is collected from pUC19 vector online and sent for synthesis to IDT.

igem#experience
None
 
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K1927000/1