Difference between revisions of "Part:BBa K4621072"

Latest revision as of 19:21, 10 October 2023

Fusion promoter composed of Pro1007 and ProP

Usage, Biology and Characterization

Pro1007 contains the promoter of GQS52_24880 in SCUT-3. The fusion promoter composed of Pro1007（BBa_K3880007）and ProP（BBa_K4621004）can control the gene expression switch while controlling its expression intensity. Usage, Biology and Characterization Pro1007 was inserted into the chitin-inducible promoter ProP to form a fusion promoter. The regulation of gene expression can be achieved by placing it before the coding of the target gene. In our study, Pro1007 stably expressed the target gene under the premise of chitin induction to control the production of ectoine and hydroxyectoine. We verified its regulation effect by LB fermentation.

For the experimenting group, we added chitin at 24 h to induce expression, and extended the total fermentation time to 5 days, to test the general accumulation.

We are surprising by the effect when combining the ProP with p24880 (p1007 & p1008), however, the results are seemingly incompatible with other data that higher expression leads to decrease on production. This phenomenon may be explained by the hypothesis of severe overexpressing leading to rapid reduction on growth and survival of SCUT-3, that p24880 carrier yield a large amount of ectoine and hydroxyectoine once induced by chitin, leading to immediate reduction in the general metabolism due to loss of living bacteria. If this case is true, p24880 is still not sufficient for long-period fermentation, and should be optimized sorely.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NotI site found at 120
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 162
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4621072 short</partinfo>
-This CRISPRi fragment contains four parts : BBa _ K4621070, BBa _ K3875019, BBa _ K4621030, and BBa _ K3081104, covering the dCas9 and sgRNA necessary for the CRISPRi system. The CRISPRi system is suitable for a variety of Streptomyces.[1] In this study, it was used to inhibit the expression of GQS52 _ 08040 gene in SCUT-3 to produce more ectoine and hydroxyectoine.
+===Usage, Biology and Characterization===
+Pro1007 contains the promoter of GQS52_24880 in SCUT-3. The fusion promoter composed of Pro1007（BBa_K3880007）and ProP（BBa_K4621004）can control the gene expression switch while controlling its expression intensity.
 Usage, Biology and Characterization
+Pro1007 was inserted into the chitin-inducible promoter ProP to form a fusion promoter. The regulation of gene expression can be achieved by placing it before the coding of the target gene. In our study, Pro1007 stably expressed the target gene under the premise of chitin induction to control the production of ectoine and hydroxyectoine. We verified its regulation effect by LB fermentation.
+For the experimenting group, we added chitin at 24 h to induce expression, and extended the total fermentation time to 5 days, to test the general accumulation.
+https://static.igem.wiki/teams/4621/wiki/parts/fermentation-with-an-inducible-promoter.png
-Due to the limited types of plasmids available for SCUT-3, we inserted the CRISPRi fragment into the previously constructed plasmid by homologous recombination to achieve simultaneous stable expression of multiple target genes. Subsequently, we verified the effectiveness of the CRISPRi system in the fermentation of ordinary LB, high-salt LB and shrimp shells.
+We are surprising by the effect when combining the ProP with p24880 (p1007 & p1008), however, the results are seemingly incompatible with other data that higher expression leads to decrease on production. This phenomenon may be explained by the hypothesis of severe overexpressing leading to rapid reduction on growth and survival of SCUT-3, that p24880 carrier yield a large amount of ectoine and hydroxyectoine once induced by chitin, leading to immediate reduction in the general metabolism due to loss of living bacteria. If this case is true, p24880 is still not sufficient for long-period fermentation, and should be optimized sorely.