Difference between revisions of "Part:BBa K2669002:Experience"

Latest revision as of 01:50, 22 October 2019

Applications of BBa_K2669002

The construct BBa_K3189015 containing the chromoprotein amilCP (BBa_K2669002) under the control of BBa_K3189001. When the system is induced with 100 ng/mL of tetracycline, a dark blue colour is produced (Figure 4 and Figure 5). This shows BBa_K3189001 is able to function with different reporter proteins other than just gfp.

Figure 1: BBa_K2669002 under the control of BBa_K3189001. amilCP expression being induced using 100 ng/mL tetracycline (left) and uninduced (right).

Figure 2: Pellets of cells of BBa_K2669002 under the control of BBa_K3189001 induced and uninduced with tetracycline. Pellet of cells induced with 100 ng/mL tetracycline (left) and pellet of cells uninduced (right).

This year iGEM Guelph was working with BBa_K2669002 as a reporter protein. When working with this chromoprotein in E.coli BL21(DE3) under the control of BBa_K3189001, we found that stronger colour was produced at lower temperatures compared to higher temperatures. In Figure 2 pigment intensity was compared between two temperatures, 25°C and 37°C. It is clear form these images that at 25°C the colonies have a darker colour compared to colonies grown at 37°C. This is evidence that AmilCP produced from BBa_K2669002 is more stable at 25°C compared to 37°C thus allowing it to build up in the cells longer before degradation resulting in darker pigmentation of the colonies.

Figure 3: BBa_K2669002 expression at 25°C and 37°C. Expression was carried out in E.coli BL21(DE3) on LB agar supplemented with 100 ug/mL ampicillin, 100 mg/mL tryptophan, 1mM IPTG, and 50 ng/mL tetracycline.

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UNIQ6f7af77689067927-partinfo-00000000-QINU UNIQ6f7af77689067927-partinfo-00000001-QINU

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 https://static.igem.org/mediawiki/parts/thumb/f/f5/T--Guelph--pTA-tetnotet.jpg/216px-T--Guelph--pTA-tetnotet.jpg
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-<b>Figure 4: [[BBa_K2669002]] under the control of [[BBa_K3189001]].</b> amilCP expression being induced using 100 ng/mL tetracycline (left) and uninduced (right).
+<b>Figure 1: [[BBa_K2669002]] under the control of [[BBa_K3189001]].</b> amilCP expression being induced using 100 ng/mL tetracycline (left) and uninduced (right).
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 https://static.igem.org/mediawiki/parts/thumb/2/29/T--Guelph--pTAtetnotetspun.jpg/207px-T--Guelph--pTAtetnotetspun.jpg
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-<b>Figure 5: Pellets of cells of [[BBa_K2669002]] under the control of [[BBa_K3189001]] induced and uninduced with tetracycline.</b> Pellet of cells induced with 100 ng/mL tetracycline (left) and pellet of cells uninduced (right).
+<b>Figure 2: Pellets of cells of [[BBa_K2669002]] under the control of [[BBa_K3189001]] induced and uninduced with tetracycline.</b> Pellet of cells induced with 100 ng/mL tetracycline (left) and pellet of cells uninduced (right).
+<br>
+<br>
+This year iGEM Guelph was working with BBa_K2669002 as a reporter protein. When working with this chromoprotein in <i>E.coli</i> BL21(DE3) under the control of [[BBa_K3189001]], we found that stronger colour was produced at lower temperatures compared to higher temperatures. In Figure 2 pigment intensity was compared between two temperatures, 25°C and 37°C. It is clear form these images that at 25°C the colonies have a darker colour compared to colonies grown at 37°C. This is evidence that AmilCP produced from [[BBa_K2669002]] is more stable at 25°C compared to 37°C thus allowing it to build up in the cells longer before degradation resulting in darker pigmentation of the colonies.
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+<br>
+https://2019.igem.org/wiki/images/thumb/f/f5/T--Guelph--AmilCPtemexpress.jpg/800px-T--Guelph--AmilCPtemexpress.jpg
+<br>
+<b>Figure 3: BBa_K2669002 expression at 25°C and 37°C.</b> Expression was carried out in <i>E.coli</i> BL21(DE3) on LB agar supplemented with 100 ug/mL ampicillin, 100 mg/mL tryptophan, 1mM IPTG, and 50 ng/mL tetracycline.
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