Difference between revisions of "Part:BBa K3889131"

 
 
(One intermediate revision by the same user not shown)
Line 4: Line 4:
  
 
BBa_B0010 Terminator check cassette for use in Bacillus subtilis.  
 
BBa_B0010 Terminator check cassette for use in Bacillus subtilis.  
 +
[[File:T--IISER-Tirupati India--Improvement2.jpeg|690px]]<br>
 +
Fig 1. Spacer replaced from <partinfo>BBa_K3889130</partinfo> by BBa_B0010
 +
 +
Formulae for terminator efficiency [1]:
 +
 +
[[File:T--IISER-Tirupati_India--Eq1.png]]
 +
 +
where
 +
 +
mCherry<sub>0</sub> &#8594; mCherry produced by device without terminator
 +
 +
sfGFP<sub>0</sub> &#8594; sfGFP produced by device without terminator
 +
 +
Using the device (<partinfo>BBa_K3889130</partinfo>) without any changes, TE<sub>Device</sub>  can be calculated which gives the expression of mCherry in absence of a terminator.
 +
 +
[[File:T--IISER-Tirupati_India--Eq2.png]]
 +
 +
where
 +
 +
mCherry &#8594; mCherry produced by the device with terminator
 +
 +
sfGFP &#8594; sfGFP produced by the device with terminator
 +
 +
and TE &#8594; is the terminator efficiency of the <partinfo>BBa_B0010</partinfo>
 +
 +
===Results===
 +
 +
[[File:T--IISER-Tirupati India--Bsfield1.png|centre|900 px]]<br>
 +
Fig. Field 1:
 +
a. B.subtilis 168 with B0010 Terminator Check Cassette-green fluoresence at wavelength 498nm (under magnification 100X)
 +
b. B.subtilis 168 with B0010 Terminator Check Cassette in bright field (under magnification 100X)
 +
c. B.subtilis 168 with B0010 Terminator Check Cassette-red fluoresence at wavelength 587nm (under magnification 100X)
 +
 +
[[File:T--IISER-Tirupati India--Bsfield2.png|900 px]]<br>
 +
Fig. Field 2:
 +
a. B.subtilis 168 with B0010 Terminator Check Cassette-green fluoresence at wavelength 498nm (under magnification 100X)
 +
b. B.subtilis 168 with B0010 Terminator Check Cassette in bright field(under magnification 100X)
 +
c. B.subtilis 168 with B0010 Terminator Check Cassette-red fluoresenceat wavelength 587nm (under magnification 100X)
 +
d. B.subtilis 168 with B0010 Terminator Check Cassette at wavelength 383nm (under magnification 100X)
 +
 +
Under a confocal microscope, we could observe the expression of sfGFP and mCherry and observe their differences in fluorescence intensities signifying that BBa_B0010 works in Bacillus subtilis.
 +
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 17: Line 59:
 
<partinfo>BBa_K3889131 parameters</partinfo>
 
<partinfo>BBa_K3889131 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
 +
===References===
 +
1. Gale, G. A. R., Wang, B., & McCormick, A. J. (2021). Evaluation and Comparison of the Efficiency of Transcription Terminators in Different Cyanobacterial Species. Frontiers in Microbiology, 11. https://doi.org/10.3389/fmicb.2020.624011

Latest revision as of 00:28, 22 October 2021


Bacillus subtilis BBa_B0010 terminator check cassette

BBa_B0010 Terminator check cassette for use in Bacillus subtilis. T--IISER-Tirupati India--Improvement2.jpeg
Fig 1. Spacer replaced from BBa_K3889130 by BBa_B0010

Formulae for terminator efficiency [1]:

T--IISER-Tirupati India--Eq1.png

where

mCherry0 → mCherry produced by device without terminator

sfGFP0 → sfGFP produced by device without terminator

Using the device (BBa_K3889130) without any changes, TEDevice can be calculated which gives the expression of mCherry in absence of a terminator.

T--IISER-Tirupati India--Eq2.png

where

mCherry → mCherry produced by the device with terminator

sfGFP → sfGFP produced by the device with terminator

and TE → is the terminator efficiency of the BBa_B0010

Results

T--IISER-Tirupati India--Bsfield1.png

Fig. Field 1: a. B.subtilis 168 with B0010 Terminator Check Cassette-green fluoresence at wavelength 498nm (under magnification 100X) b. B.subtilis 168 with B0010 Terminator Check Cassette in bright field (under magnification 100X) c. B.subtilis 168 with B0010 Terminator Check Cassette-red fluoresence at wavelength 587nm (under magnification 100X)

T--IISER-Tirupati India--Bsfield2.png
Fig. Field 2: a. B.subtilis 168 with B0010 Terminator Check Cassette-green fluoresence at wavelength 498nm (under magnification 100X) b. B.subtilis 168 with B0010 Terminator Check Cassette in bright field(under magnification 100X) c. B.subtilis 168 with B0010 Terminator Check Cassette-red fluoresenceat wavelength 587nm (under magnification 100X) d. B.subtilis 168 with B0010 Terminator Check Cassette at wavelength 383nm (under magnification 100X)

Under a confocal microscope, we could observe the expression of sfGFP and mCherry and observe their differences in fluorescence intensities signifying that BBa_B0010 works in Bacillus subtilis.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 121


References

1. Gale, G. A. R., Wang, B., & McCormick, A. J. (2021). Evaluation and Comparison of the Efficiency of Transcription Terminators in Different Cyanobacterial Species. Frontiers in Microbiology, 11. https://doi.org/10.3389/fmicb.2020.624011