Difference between revisions of "Part:BBa K3183010"

Revision as of 01:19, 22 October 2019

mClover3 Fluorescent Protein, Codon Optimized for L. reuteri

mClover3 is a green fluorescent protein derivative which has been codon optimized for Lactobacillus reuteri 10023C, and may have uses in other Lactobacillus species.

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
COMPATIBLE WITH RFC[1000]

Fluorescence wavelengths

Bajar et al report the following excitation and emission data for mClover3 -

Excitation max - 506nm
Emission max - 518nm

Figure 1: mClover3 spectrum

Parts characterized by Oxford iGEM 2019

This part was characterised in the composite part BBa_K3183028, and BBa_K3183101.

Additionally, with the purified protein (BBa_K3183203), we were able to make a standard curve for mClover3 with a fluorometer. We used two different buffers: phosphate buffered saline (PBS) and De Man, Rogosa and Sharpe (MRS) media.

Figure 1: mClover3 standard curve - Fluorescence intensity vs Concentration (mg/ml). On the graph, we can observe that in MRS the FI is lower than that in PBS. This could be due to the high fluorescence of MRS that masks mClover3 fluorescence.Error bars represent 1 s.d., n=3

Figure 2: - ldh promoter FI and OD600 time dependence - Blank corrected Fluorescence intensity and OD600 was plotted against time for ldh promoter. A large peak in OD600 can be observed, which could be an outliar due to random error in the instrument. Error bars represent Standard error of the mean. n=3

Use by Team Oxford 2019

This part was used in the following composite parts: BBa_K3183028, BBa_K3183300, BBa_K3183104, BBa_K3183203, BBa_K3183101, and BBa_K3183104.

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 Additionally, with the purified protein (<partinfo>BBa_K3183203</partinfo>), we were able to make a standard curve for mClover3 with a fluorometer. We used two different buffers: phosphate buffered saline (PBS) and De Man, Rogosa and Sharpe (MRS) media.
-[[File:BBa_K3183028_mClover3_log_standardcurve.png|thumb|left|430px|'''Figure 1:'''  mClover3 standard curve - Fluorescence intensity vs Concentration (mg/ml). On the graph, we can observe that in MRS the FI is lower than that in PBS. This could be due to the high fluorescence of MRS that masks mClover3 fluorescence.<i>Error bars represent 1 s.d., n=3</i>]]
+[[File:BBa_K3183028_mClover3_log_standardcurve.png|thumb|center|430px|'''Figure 1:'''  mClover3 standard curve - Fluorescence intensity vs Concentration (mg/ml). On the graph, we can observe that in MRS the FI is lower than that in PBS. This could be due to the high fluorescence of MRS that masks mClover3 fluorescence.<i>Error bars represent 1 s.d., n=3</i>]]
-[[File:T--Oxford--Results-ldh.png|thumb|right|430px|'''Figure 2:''' - ldh promoter FI and OD600 time dependence - Blank corrected Fluorescence intensity and OD600 was plotted against time for ldh promoter. A large peak in OD600 can be observed, which could be an outliar due to random error in the instrument. <i> Error bars represent Standard error of the mean. n=3 </i>]]
+[[File:T--Oxford--Results-ldh.png|thumb|center|430px|'''Figure 2:''' - ldh promoter FI and OD600 time dependence - Blank corrected Fluorescence intensity and OD600 was plotted against time for ldh promoter. A large peak in OD600 can be observed, which could be an outliar due to random error in the instrument. <i> Error bars represent Standard error of the mean. n=3 </i>]]
 <br><br>
 ===Use by Team Oxford 2019===
 This part was used in the following composite parts: <partinfo>BBa_K3183028</partinfo>, <partinfo>BBa_K3183300</partinfo>, <partinfo>BBa_K3183104</partinfo>, <partinfo>BBa_K3183203</partinfo>, <partinfo>BBa_K3183101</partinfo>, and <partinfo>BBa_K3183104</partinfo>.