Difference between revisions of "Part:BBa K1648004"
Fatfaatlai (Talk | contribs) |
Fatfaatlai (Talk | contribs) |
||
(2 intermediate revisions by the same user not shown) | |||
Line 10: | Line 10: | ||
A GFP-nanobody([https://parts.igem.org/Part:BBa_K1648005 K1648005]) is added to this part in order to express GFP-nanobody on the magnetosome. This new part is [https://parts.igem.org/Part:BBa_K1648006 K1648006] | A GFP-nanobody([https://parts.igem.org/Part:BBa_K1648005 K1648005]) is added to this part in order to express GFP-nanobody on the magnetosome. This new part is [https://parts.igem.org/Part:BBa_K1648006 K1648006] | ||
− | This Part were verified by double digestion (Fig. 1). | + | This Part were verified by double digestion (Fig. 1) and sequencing. |
[[File:K1648004gel.jpg]] | [[File:K1648004gel.jpg]] | ||
− | Fig 1. The photo of 1% agarose gel electrophoresis showing PCR products | + | Fig 1. The photo of 1% agarose gel electrophoresis showing PCR products. (A) L: DNA ladder. Lane 1: PCR products of linear Insertion Kit. |
Latest revision as of 13:58, 25 September 2015
Insertion Kit for Fusing Protein of Interest to Magnetosome Membrane
Consist of a gene (mamC) from magnetosome island of Magnetospirillum gryphiswaldense genome, a promotor & RBS(J13002) in front of multiple restriction sites and a double terminator(B0015) at the back of multiple restriction sites.
Different genes can be added into this part at multiple restriction sites, so that different proteins will be expressed on the magnetosome after transformation of the part into A. vinelandii genome via random integration.
A GFP-nanobody(K1648005) is added to this part in order to express GFP-nanobody on the magnetosome. This new part is K1648006
This Part were verified by double digestion (Fig. 1) and sequencing.
Fig 1. The photo of 1% agarose gel electrophoresis showing PCR products. (A) L: DNA ladder. Lane 1: PCR products of linear Insertion Kit.
RFP is inserted in Insertion Kit for biobrick standard. The Insertion kit with RFP is transformed in E. coli (DH5 alpha). Expression of RFP in Insertion is shown(Fig 2.).
Fig 2. Insertion kit with RFP(BBa_K1648004) is expressed in E. coli (DH5 alpha)
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2759
Illegal XbaI site found at 2765
Illegal SpeI site found at 1
Illegal PstI site found at 7 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2759
Illegal SpeI site found at 1
Illegal PstI site found at 7 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2759
Illegal XhoI site found at 1162
Illegal XhoI site found at 2188 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2759
Illegal XbaI site found at 2765
Illegal SpeI site found at 1
Illegal PstI site found at 7 - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2759
Illegal XbaI site found at 2765
Illegal SpeI site found at 1
Illegal PstI site found at 7
Illegal NgoMIV site found at 2546
Illegal NgoMIV site found at 2636 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 2611