Part:BBa_K3037003

Fusion protein dCas9 + HRP (MBP/dCas9/linker/HRP/Strep-tag)

Fusion protein
Function	Colour detection of specific DNA sequences
Use in	Escherichia coli
RFC standard	RFC 25 compatible
Backbone	pSB1C3
Submitted by	Team:TU_Dresden 2019[1]

Overview

The TU Dresden 2019 team design this biobrick in order to create a system for the short and quick detection of specific DNA sequences.

The plasmid used for the expression of the construct was pSB1C3.

Characterization

Outline

We performed the following characterization experiments:

1) Expression in pOOC97 (BBa_K3037000):

Experiments in Detail

1) Expression in pOOC97 (BBa_K3037000):

The fusion protein was expressed using as a backbone the plasmid pOOC97 (BB_aK3037000)

The purpose of this experiment is to show that the Escherichia coli grows normaly after the induction of the expression of the fusion protein.

For this the development of the culture was monitored by measuring the OD at 600 nm during different time before and after induction with 1 mM IPTG. As shown in the curve the growing of the bacteria is not affected by the expression of the protein.

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Sequence

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 2302
  Illegal NheI site found at 5500
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 381
  Illegal BglII site found at 5742
  Illegal BamHI site found at 4581
  Illegal BamHI site found at 5314
  Illegal XhoI site found at 5826
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 79