Difference between revisions of "Part:BBa K1648000"
Fatfaatlai (Talk | contribs) |
Fatfaatlai (Talk | contribs) |
||
(5 intermediate revisions by the same user not shown) | |||
Line 2: | Line 2: | ||
<partinfo>BBa_K1648000 short</partinfo> | <partinfo>BBa_K1648000 short</partinfo> | ||
[[File:K1648000_Map.png|350px|thumb|right|Construct Map for K1648000]] | [[File:K1648000_Map.png|350px|thumb|right|Construct Map for K1648000]] | ||
− | |||
− | |||
− | |||
<div style="text-align:justify; text-justify:inter-ideograph"> | <div style="text-align:justify; text-justify:inter-ideograph"> | ||
Line 12: | Line 9: | ||
Due to potential difficulty in introducing the large magnetosome formation operons into ''Azotobacter vinelandii'', a "template" consists of flanking sequences from '''mamAB operon''' is designed for '''homologous recombination'''. | Due to potential difficulty in introducing the large magnetosome formation operons into ''Azotobacter vinelandii'', a "template" consists of flanking sequences from '''mamAB operon''' is designed for '''homologous recombination'''. | ||
− | Part containing this template ([https://parts.igem.org/Part:BBa_K1648002 K1648002]) is introduced into ''A. vinelandii'' genome via '''random integration''' before full operon sequences to be transformed into ''A. vinelandii''. | + | Part containing this template ([https://parts.igem.org/Part:BBa_K1648002 K1648002]) is introduced into ''A. vinelandii'' genome via '''random integration''' before full operon sequences to be transformed into ''A. vinelandii''. This Part were verified by double digestion (Fig. 1) and sequencing. |
− | + | ||
− | + | [[File:K16480000.jpg|400px]] | |
+ | |||
+ | Fig. 1 Checking of recombinant plasmid using double digestion. | ||
L: DNA ladder. | L: DNA ladder. | ||
− | Lane 1-3: Recombination Template for mamAB Operon (BBa_K1648000) without digestion, with single digestion at XbaI site, with double digestion cut at XbaI and PstI site. | + | Lane 1-3: Recombination Template for mamAB Operon (BBa_K1648000) without digestion, with single digestion at XbaI site, with double digestion cut at XbaI and PstI site. |
− | + | ||
</div> | </div> | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Latest revision as of 23:07, 24 September 2015
Recombination Template for mamAB Operon
Consists of four fragments from magnetosome island of Magnetospirillum gryphiswaldense genome.
Due to potential difficulty in introducing the large magnetosome formation operons into Azotobacter vinelandii, a "template" consists of flanking sequences from mamAB operon is designed for homologous recombination.
Part containing this template (K1648002) is introduced into A. vinelandii genome via random integration before full operon sequences to be transformed into A. vinelandii. This Part were verified by double digestion (Fig. 1) and sequencing.
Fig. 1 Checking of recombinant plasmid using double digestion. L: DNA ladder. Lane 1-3: Recombination Template for mamAB Operon (BBa_K1648000) without digestion, with single digestion at XbaI site, with double digestion cut at XbaI and PstI site.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 775
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 291
Illegal AgeI site found at 810 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 536