Difference between revisions of "Part:BBa K2912015"

(→‎Usage and Biology)
(→‎Usage and Biology)
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===Usage and Biology===
 
===Usage and Biology===
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The lysis gene from colicin-producing strains of bacteria encodes the lysis protein which can cause the host cell to lyze as long as the protein is activated. On the basic of its biological function, we can use lysis gene to break our engineering E.coli and release the  interior cell contents when it's time to extract our product-shRNA by adding a Tryptophan attenuator between a RBS and lysis gene
+
The lysis gene from colicin-producing strains of bacteria encodes the lysis protein which can cause the host cell to lyze as long as the protein is activated. On the basic of its biological function, we can use lysis gene to break our engineering E.coli and release the  interior cell contents when it's time to extract our product-shRNA by adding a Tryptophan attenuator between a RBS and lysis gene.
  
−
We transformed the expression vector into HT115 (DE3) E.coli by heat-shock method, and they were cultured in 1 LB liquid medium to exact exponential phase. Afterwards, we started a cell growth curve experiment of our engineering E.coli(illustrated with Fig.1)
+
Before the experiment of our part-Trp_Lysis gene. We have done the T7_lysis gene charaterization experiment by IPTG inducing in LB solid medium(illustrated with Fig.2).
 +
 
 +
According to the charaterization experiment's result, we can draw a conclusion that once the lysis protein is produced by IPTG inducing, their growth will be significantly inhibited.
 +
 +
We transformed the expression vector into HT115 (DE3) E.coli by heat-shock method, and they were cultured in 1 LB liquid medium to exact exponential phase. Afterwards, we started a cell growth curve experiment of our engineering E.coli in different tryptophan concentration gradients (illustrated with Fig.3).
 +
 
 +
Compared to T7 promotor-lysis, the Trp_Lysis gene obiviously has the advantage which is able to kill the host cell by breaking their structure in a controlled way. In other words, we can swich off or swich on the killing metabolism by controlling the tryptophan concentration of the culture medium, and the concentration critical valve is around 0.3%(if the concentration is more than 0.3%, the killing metabolsim will be closed so that the host cells can grow as usual)
  
  

Revision as of 02:12, 17 October 2019


Trp_Lysis gene

Trp_Lysis gene expression can be controlled by the concentration of Tryptophan

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage and Biology

The lysis gene from colicin-producing strains of bacteria encodes the lysis protein which can cause the host cell to lyze as long as the protein is activated. On the basic of its biological function, we can use lysis gene to break our engineering E.coli and release the interior cell contents when it's time to extract our product-shRNA by adding a Tryptophan attenuator between a RBS and lysis gene.

Before the experiment of our part-Trp_Lysis gene. We have done the T7_lysis gene charaterization experiment by IPTG inducing in LB solid medium(illustrated with Fig.2).

According to the charaterization experiment's result, we can draw a conclusion that once the lysis protein is produced by IPTG inducing, their growth will be significantly inhibited.

We transformed the expression vector into HT115 (DE3) E.coli by heat-shock method, and they were cultured in 1 LB liquid medium to exact exponential phase. Afterwards, we started a cell growth curve experiment of our engineering E.coli in different tryptophan concentration gradients (illustrated with Fig.3). 

Compared to T7 promotor-lysis, the Trp_Lysis gene obiviously has the advantage which is able to kill the host cell by breaking their structure in a controlled way. In other words, we can swich off or swich on the killing metabolism by controlling the tryptophan concentration of the culture medium, and the concentration critical valve is around 0.3%(if the concentration is more than 0.3%, the killing metabolsim will be closed so that the host cells can grow as usual)