Team:UMaryland/project/results

From 2014.igem.org

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<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 5 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing Results and Chromatogram of SDM’d OmpA in pSB1C3</span></p>
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 5 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing Results and Chromatogram of SDM’d OmpA in pSB1C3</span></p>
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<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><br /></span><span style="line-height: 12.777777671814px;">"The previous version of Lpp-OmpA-Linker (BBa_K103006), designed for the pSB1A2 vector backbone, contained the non-traditional restriction sites NdeI (CATATG) and SacI (GAGCTC) in order to attach proteins at the N-terminus and C-terminus of OmpA, respectively. While the NdeI site is still useful for RE cloning at the N-terminus of OmpA, the implementation of the new plasmid backbone pSB1C3 has made the SacI site difficult to use. This is due to the presence of a second SacI site in the chloramphenicol acetyltransferase gene in pSB1C3. We have thus decided to mutagenize the previous SacI site on Lpp-OmpA-Linker into a KasI site in order to remove this issue. The KasI site (GGCGCC) encodes the amino acids glycine and alanine, converting the sequence of the linker from Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ser to Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ala. We believe this to be a sufficiently minor alteration to the unfolded linker sequence as to not affect protein function."</span></p>  
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<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><br /></span><span style="line-height: 12.777777671814px;">The previous version of Lpp-OmpA-Linker (BBa_K103006), designed for the pSB1A2 vector backbone, contained the non-traditional restriction sites NdeI (CATATG) and SacI (GAGCTC) in order to attach proteins at the N-terminus and C-terminus of OmpA, respectively. While the NdeI site is still useful for RE cloning at the N-terminus of OmpA, the implementation of the new plasmid backbone pSB1C3 has made the SacI site difficult to use. This is due to the presence of a second SacI site in the chloramphenicol acetyltransferase gene in pSB1C3. We have thus decided to mutagenize the previous SacI site on Lpp-OmpA-Linker into a KasI site in order to remove this issue. The KasI site (GGCGCC) encodes the amino acids glycine and alanine, converting the sequence of the linker from Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ser to Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ala. We believe this to be a sufficiently minor alteration to the unfolded linker sequence as to not affect protein function.</span></p>  

Revision as of 20:45, 16 October 2014

Results

About Umaryland

UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!