Difference between revisions of "Part:BBa K2559008"

Revision as of 08:31, 15 October 2018

Neutral site for homologous recombination

Neutral site cloned from Synechocystis sp. PCC 6803, the key component of gene expression platform

Usage and Biology

Foreign genes and even entire pathways are often ported into chassis organisms, requiring either plasmidbased expression or identification of a neutral site for genome integration. As genome integration is more stable and predictable compared to plasmid based expression,this is often the preferred method for modification,particularly for industrial microbial strains.[1] Nowadays several putative neutral integration sites have been identified in recent efforts, including the pseudogene glpK (SYNPCC7002_A2842) [2] and the genomic region between hypothetical protein genes (SYNPCC7002_A0935 and SYNPCC7002_A0936) [3], yet the neutrality of these sites remains to be verified. Liang[4] have successfully designed and constructed using synthetic biology approach two recombinant strainsof Synechococcus elongatus PCC7942 for heterologous expression of different industriallyrelevant Bacillus subtilis CotA laccase. Of the two strains the one with CotA laccase integrated in neutral site II(PCC7942-NSII-CotA) was superior in terms of growth rate and enzymatic activity towardtypical laccase substrates: ABTS [2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonate)] andsyringaldazine. That may suggest that two of the traditionally used neutral sites ofS. elongatus PCC7942 are not equally suitable for the expression of certain transgenes

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 1326
Illegal BamHI site found at 1537
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 661
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 776
Illegal BsaI.rc site found at 1886

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2559008 short</partinfo>
-It allows us to use the culture temperature to regulate the expression of downstream genes
+Neutral site cloned from Synechocystis sp. PCC 6803, the key component of gene expression platform
 ===Usage and Biology===
-The promoter of HSPs is a strong promoter, the amount of the bicistronic, 2.3 kb transcript of groESL1 operon increased 30-fold within 30 min upon heat shock. It allows us to regulate the expression of target genes by changing the culture temperature, and avoid the risk of losing transformed algae strains for toxic effect caused by target genes.
+Foreign genes and even entire pathways are often ported into chassis organisms, requiring either plasmidbased expression or identification of a neutral site for genome integration. As genome integration is more stable and predictable compared to plasmid based expression,this is often the preferred method for modification,particularly for industrial microbial strains.[1]
+Nowadays several putative neutral integration sites have been identified in recent efforts, including the pseudogene glpK (SYNPCC7002_A2842) [2] and the genomic region between hypothetical protein genes (SYNPCC7002_A0935 and SYNPCC7002_A0936) [3], yet the neutrality of these sites remains to be verified.
+Liang[4] have successfully designed and constructed using synthetic biology approach two recombinant strainsof Synechococcus elongatus PCC7942 for heterologous expression of different industriallyrelevant Bacillus subtilis CotA laccase. Of the two strains the one with CotA laccase integrated in neutral site II(PCC7942-NSII-CotA) was superior in terms of growth rate and enzymatic activity towardtypical laccase substrates: ABTS [2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonate)] andsyringaldazine. That may suggest that two of the traditionally used neutral sites ofS. elongatus PCC7942 are not equally suitable for the expression of certain transgenes
-<!-- Add more about the biology of this part here
-===Usage and Biology===
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