Difference between revisions of "Part:BBa K510022"

Latest revision as of 22:41, 24 September 2011

attTn7

Single-target transposition to the bacterial genomes is dependent on the presence of a conserved sequence, encompasing the 3' end of the glmS gene, encoding glucosamine synthetase, and part the intergenic region immediately downstream, designated the attTn7 site (from attachment site). Our [http://2011.igem.org/Team:UPO-Sevilla/Foundational_Advances/MiniTn7/Bioinformatics/attTn7_Insertion_Site Bioinformatic analysis] reveal that this region is highly conserved, and Tn7 transposition has been demonstrated in over 20 bacterial species ([http://www.ncbi.nlm.nih.gov/pubmed/8556868 Craig, 1996]). However, the lack of a suitable target may be the limiting factor in some of the organisms in which Tn7 transposition does not occur, including a variety of bacteria, but also archea and eukaryotes. The possibility of inserting a functional attTn7 in the genomes of these organisms may enable site-specific Tn7 transposition for at least some of them, thus expanding the host range of this transposon and its derived tools, such as the [http://2011.igem.org/Team:UPO-Sevilla/Foundational_Advances/MiniTn7/Overview miniTn7 BioBrick toolkit] (BBa_K510000, BBa_K510012).

Figure. The organization of attTn7 (from [http://www.mobilednajournal.com/content/1/1/18 Rupak et al, 2010]). A schematic of the attTn7 at the C-terminus of the glmS gene with the TnsD binding site is shown. The sequence of the E. coli GlmS protein, and the consensus attTn7 sequence were derived as described in the text (note that the least conserved nucleotides correspond to the third position of each codon).

More information about this plasmid from the UPO-Sevilla 2011 team in the Experience section.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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-TnsABC+D transposition machinery causes the site-specific insertion of the Tn7 transposon in the attTn7 (from attachment site) sequence, which is found in many bacteria. Our [http://2011.igem.org/Team:UPO-Sevilla/Foundational_Advances/MiniTn7/Bioinformatics/attTn7_Insertion_Site Bioinformatic analysis] reveal that the insertion site is well conserved in Gram-negative bacteria, an even in higher organisms until humans. However, here we add a portable attTn7 to allow using the [http://2011.igem.org/Team:UPO-Sevilla/Foundational_Advances/MiniTn7/Overview miniTn7 BioBrick toolkit] in any organism as well as in plasmids.
+'''Single-target transposition to the bacterial genomes''' is dependent on the presence of a conserved sequence, encompasing the 3' end of the glmS gene, encoding glucosamine synthetase, and part the intergenic region immediately downstream, designated the '''attTn7 site''' (from attachment site). Our [http://2011.igem.org/Team:UPO-Sevilla/Foundational_Advances/MiniTn7/Bioinformatics/attTn7_Insertion_Site Bioinformatic analysis] reveal that this region is highly conserved, and Tn7 transposition has been demonstrated in over 20 bacterial species ([http://www.ncbi.nlm.nih.gov/pubmed/8556868 Craig, 1996]). However, the lack of a suitable target may be the limiting factor in some of the organisms in which Tn7 transposition does not occur, including a variety of bacteria, but also archea and eukaryotes. The possibility of inserting a functional attTn7 in the genomes of these organisms may enable site-specific Tn7 transposition for at least some of them, thus expanding the host range of this transposon and its derived tools, such as the [http://2011.igem.org/Team:UPO-Sevilla/Foundational_Advances/MiniTn7/Overview miniTn7 BioBrick toolkit] ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K510000 BBa_K510000], [https://parts.igem.org/wiki/index.php?title=Part:BBa_K510012 BBa_K510012]).
-All the derivatives of the miniTn7 BioBrick toolkit could be inserted in the attTn7 site by transposition.
+[[Image:UPOSevilla-attTn7-glmS.jpg|700px|center]]
+'''Figure. The organization of attTn7''' (from [http://www.mobilednajournal.com/content/1/1/18 Rupak et al, 2010]). A schematic of the attTn7 at the C-terminus of the glmS gene with the TnsD binding site is shown. The sequence of the E. coli GlmS protein, and the consensus attTn7 sequence were derived as described in the text (note that the least conserved nucleotides correspond to the third position of each codon).
+More information about this plasmid from the UPO-Sevilla 2011 team in the ''Experience section''.
 <!-- Add more about the biology of this part here