Team:Imperial College/Growth Curve

From 2008.igem.org

(Difference between revisions)
Line 13: Line 13:
{{Imperial/Box1|The Model|
{{Imperial/Box1|The Model|
-
The model illustrates the main growth phases the ''B. subtilis'' undergoes.  These are identified as the '''lag phase''', the '''exponential phase''' and the '''stationary phase'''.  The death phase is a constitutive event and it is possible that it exerts an influence on the three phases discussed below.  However, to simplify a complicated model, it is less relevant in this case and therefore is not included in this model.
+
Our model is based on the crucial role played by nutrients ,and other similar resources, during the main growth phases of B. subtilis undergoes - the '''lag phase''', the '''exponential phase''' and the '''stationary phase'''.
[[Image:Lag_phase.JPG|300px|left]]
[[Image:Lag_phase.JPG|300px|left]]

Revision as of 19:55, 29 October 2008


Modelling the Growth Curve

The first task of our dry lab team was to construct a simple yet effective model for the growth curve of B. subtilis.
The inspiration for our model came from the work done by the Imperial College team on the characterisation of Cell Free Systems [iGEM2007] and the elegant manner they found to couple the production terms in the model with the resources available in the medium. .




The Model


Our model is based on the crucial role played by nutrients ,and other similar resources, during the main growth phases of B. subtilis undergoes - the lag phase, the exponential phase and the stationary phase.

Lag phase.JPG
Lag Phase

During the lag phase, the rate of growth is slow. All nutrients are situated outside the cell initially. Some time is needed for an adequate amount of nutrients to move from the outside of the cell into the interior of the cell. This is vital as the cell requires the nutrients for growth.






Exponential phase.JPG
Exponential Phase

Nutrients are consumed during the cell growth and the growth is exponential as long as there are enough nutrients available. The exchange of nutrients ensures that the intra- and extracellular nutrient concentration are the same.







Stationary phase2.JPG
Stationary Phase

The growth of the colony ceases in number and in volume. This happens when the colony has consumed all available nutrients. Other contributing factors may be death and cell division.









The growth is represented in terms of volume. By doing so we can, to some extent, overlook the process of cell division during the bacteria growth. We can assume the internal concentration of nutrients is reset almost instantaneously to the external concentration. During this time, the total volume does not change.

The growth of the overall volume V is therefore modelled as:

Growth equ.JPG

The M-file used to generate the model below is located in the Appendices section of the Dry Lab hub.

Below is a typical model used for modelling the growth of bacteria. A high growth rate, as illustrated by the model below, will cause nutrients to be uptaken quickly resulting in a 'nutrient crash'. As the amount of resources are limited, the growth curve eventually become flat in the stationary phase. This is when the growth of the bacteria ceases.

Nutrient ft.JPGLabel model.JPG

Results


The model for the growth curve was fitted to the experimental results. The results are shown below. The experimental results are depicted by the red curve, while our model is shown by the green curve. The resource curve was also plotted as a function of time and is shown below.

The model 'fit' is very good in the exponential and stationary phases. However, this does not apply to the lag phase. This suggests that the model should be expanded in order to take into account phenomena such as the movement of the nutrients into the cells.


Experimental Result.JPG
Fitted Curve.JPG
Resource Curve.JPG
Experimental Results
Fitted Curve
Resource Curve



The search for the optimal model parameters was done by brute force. But instead of browsing the whole space of parameters, we reduced the search volume with a simple pre-processing of the experimental data. The initial volume was estimated from the data - likewise the volume in the stationary phase which is directly related to the model parameters R0 and alpha. The model parameter A being related to the apparent growth rate of the experimental data, We plotted the log graph to determine the growth rate of our data.

Log-Graph used to determine the growth rate

The following constants were found to yield the best fit to experimental results:
- GROWTH CONSTANT (A): 1.3494
- INITIAL NUTRIENT CONCENTRATION (R0): 2
- HILL COEFFICIENT (n): 1.25
- INITIAL OD: 0.4
- CONSTANT (α): 0.64516