Part:BBa_K5048065:Experience

Experiments

This part acts as a system that provides more protein HSP60 in the supernatant.

The overall experiments we conducted on BBa_K5048065 include the following:

1. Verification of PCR products;
2. Verification of transformation of the plasmid.

Results

Verification of PCR products

For the genetic circuit SUMO-HSP60, DNA segments, including SUMO and HSP-pET-backbone, were amplified successfully (Fig 1).

Fig 1 The construction of SUMO-HSP60
A. The agarose gel electrophoresis of SUMO segment for SUMO-HSP60. SUMO: 325bp.
B. The agarose gel electrophoresis of HSP60-pET-backbone segment for SUMO-HSP60. HSP60-pET-backbone: 6982bp.

Verification of transformation of the plasmid

Furthermore, we connected the two DNA segments and built up plasmid SUMO-HSP60 in E. coli BL21(DE3) with single colonies containing the target band (Fig 2).

Fig 1 SUMO-HSP60 successfully transferred into E. coli BL21(DE3)
A. The plate of E. coli DH5α harboring the transferred plasmid SUMO-HSP60, which contains abundant single colonies. Resistance: Kanamycin (Kan).
B. The colony PCR for E. coli BL21(DE3). SUMO-HSP60 1 and 3-5 all have segments corresponding to the estimated DNA length. SUMO-HSP60: 2055bp
C. The sequencing result for SUMO-HSP60 in E. coli BL21(DE3), demonstrates the successful transformation.

Applications of BBa_K5048065

This part acts as a system that provides more protein HSP60 in the supernatant to achieve better results for Co-IP.

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