Difference between revisions of "Part:BBa K3963001"

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==Experiments and results==
 
==Experiments and results==
  
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===Plasmid===
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===Plasmid characterization===
  
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The plasmid should have a size of 5560 bp but in the gel the plasmid looks smaller about 4 kb (see Fig. 2A).We transformed the plasmid pBAV1k-lacI-Trc-beta-agarase YM01-3 into <I>E. coli</I> DH5ɑ and the typical pit formation can be observed (see Fig. 2B). We send the plasmid to seuquencing to control the beta-agarase insert (see Fig. 2B).
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The plasmid should have a size of 5560 bp but in the gel the plasmid looks smaller about 4 kb (see Fig. 2A).We transformed the plasmid pBAV1k-lacI-Trc-beta-agarase YM01-3 into <I>E. coli</I> DH5ɑ and the typical pit formation can be observed (see Fig. 2B). We send the plasmid to sequencing to control the beta-agarase insert (see Fig. 2C).
  
 
Figure 2
 
Figure 2

Revision as of 13:38, 19 October 2021


pBAV1k-lacI-Trc-beta-agarase YM01-3

The plasmid is combined by the backbone from the pBWB162 plasmid (BBa_K3963003) and cloned by restriction based cloning with the insert beta-agarase YM01-3 (BBa_K2094002). mCherry protein was cut out. The plasmid still contains the lacI behind lacIq promoter and a kanamycin selection marker.

Design

Figure 1 Plasmid pBAV1k-lacI-Trc-beta-agarase YM01-3 design. The plasmid size is 5560 bp.

Experiments and results

Plasmid characterization

The plasmid should have a size of 5560 bp but in the gel the plasmid looks smaller about 4 kb (see Fig. 2A).We transformed the plasmid pBAV1k-lacI-Trc-beta-agarase YM01-3 into E. coli DH5ɑ and the typical pit formation can be observed (see Fig. 2B). We send the plasmid to sequencing to control the beta-agarase insert (see Fig. 2C).

Figure 2

Natural transformation function

In Figure 3 it can be seen that after cloning of the pBWB162 plasmid (BBa_K3963000) to the recent plasmid the ability to be taken up by Acinetobacter baylyi ADP1, a natural competent bacterial strain, is still existing.

Figure 3

Discussion

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 2292
    Illegal BamHI site found at 3167
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 3183
    Illegal AgeI site found at 3220
  • 1000
    COMPATIBLE WITH RFC[1000]