Difference between revisions of "Part:BBa K3748015"

Revision as of 10:47, 11 October 2022

pREV1

Weak strenght yeast promoter S.cerevisiae.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 705
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Team Estonia_TUIT characterization of BBa_K3748015 (pREV1)

pREV1 is a constitutive promoter responsible for the expression of REV1, a gene encoding bi-functional DNA-directed DNA polymerase/deoxycytidyl transferase (Lee et al., 2015). This enzyme is involved in error-prone translesion synthesis, one of the pathways for DNA repair (Lawrence, 2004).

We quantified the expression driven by pREV1 using fluorescent proteins as reporter genes. The expression cassettes were integrated into the yeast genome, and the fluorescence of the reporter proteins was monitored by quantitative time-lapse microscopy. The fluorescence levels were compared to the cell background fluorescence of the control DOM90 strain, which does not express any fluorescent proteins

pREV1 promoters showed a constitutive level of Venus fluorescence throughout the experiment, confirming that it is a constitutive promoter. Compared to the background fluorescence of the DOM90 strain pREV1 demonstrated a five-fold higher level of fluorescence intensity (Fig. 1).

Figure 1. The expression level of Venus is controlled by pREV1. Bars indicate the mean fluorescence signal (AU), and error bars show the standard deviation.

References:

Lee, M. E., DeLoache, W. C., Cervantes, B., & Dueber, J. E. (2015). A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly. ACS Synthetic Biology, 4(9), 975–986. https://doi.org/10.1021/SB500366V/SUPPL_FILE/SB500366V_SI_002.ZIP
Lawrence, C. W. (2004). Cellular functions of DNA polymerase ζ and REV1 protein. Advances in Protein Chemistry, 69, 167–203. https://doi.org/10.1016/S0065-3233(04)69006-1

@@ Line 19: / Line 19: @@
 ==Team Estonia_TUIT characterization of BBa_K3748015 (<i>pREV1</i>)==
-pREV1 is a constitutive promoter responsible for the expression of REV1, a gene encoding bi-functional DNA-directed DNA polymerase/deoxycytidyl transferase (Lee <i>et al</i>., 2015). This enzyme is involved in error-prone translesion synthesis, one of the pathways for DNA repair (Lawrence, 2004).
+<i>pREV1</i> is a constitutive promoter responsible for the expression of <i>REV1</i>, a gene encoding bi-functional DNA-directed DNA polymerase/deoxycytidyl transferase (Lee <i>et al</i>., 2015). This enzyme is involved in error-prone translesion synthesis, one of the pathways for DNA repair (Lawrence, 2004).
-We quantified the expression driven by REV1 using fluorescent proteins as reporter genes. The expression cassettes were integrated into the yeast genome, and the fluorescence of the reporter proteins was monitored by quantitative time-lapse microscopy. The fluorescence levels were compared to the cell background fluorescence of the control DOM90 strain, which does not express any fluorescent proteins
+We quantified the expression driven by <i>pREV1</i> using fluorescent proteins as reporter genes. The expression cassettes were integrated into the yeast genome, and the fluorescence of the reporter proteins was monitored by quantitative time-lapse microscopy. The fluorescence levels were compared to the cell background fluorescence of the control DOM90 strain, which does not express any fluorescent proteins
-pREV1 promoters showed a constitutive level of Venus fluorescence throughout the experiment, confirming that it is a constitutive promoter. Compared to the background fluorescence of the DOM90 strain pREV1 demonstrated a five-fold higher level of fluorescence intensity (Fig. 1).
+<i>pREV1</i> promoters showed a constitutive level of Venus fluorescence throughout the experiment, confirming that it is a constitutive promoter. Compared to the background fluorescence of the DOM90 strain <i>pREV1</i> demonstrated a five-fold higher level of fluorescence intensity (Fig. 1).
-[[Image:PREV1 Estonia TUIT new.png|600px|thumb|center|<b>Figure 1. The expression level of Venus is controlled by pREV1</b>. Bars indicate the mean fluorescence signal (AU), and error bars show the standard deviation. ]]
+[[Image:PREV1 Estonia TUIT new.png|600px|thumb|center|<b>Figure 1. The expression level of Venus is controlled by <i>pREV1</i></b>. Bars indicate the mean fluorescence signal (AU), and error bars show the standard deviation. ]]
 <b>References:</b>