Part:BBa_K2559008
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1326
Illegal BamHI site found at 1537 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 661
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 776
Illegal BsaI.rc site found at 1886
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