From 2007.igem.org

Results of SDS-PAGE Gel of Soluble Proteins

Invitrogen BenchMark Protein Ladder

Plasmid Key for Protein Gels

Discussion

Prior to running the protein gels:

*We expected to see protein bands for the constructs that contained:

1. -H gene (~140kDa)that were induced with IPTG

2. -I gene(~38kDa) that were induced with IPTG

*We did not expect to see bands that coded for:

1. -D gene (~70kDa)

2. - All constructs that were not induced with IPTG

• Although the induced constructs that contained the -H gene had strong bands at ~140kDa, the uninduced constructs also showed strong bands in the same area. The -H gene was able to be expressed with or without induction of IPTG.

• In addition, protein bands that coded for the -I gene showed a strong band, when alone.

• However, when the -I gene was expressed with the -H gene, it did not clearly show a strong band at ~38 kDa in comparison to the benchmark ladder.

• As expected, protein bands for the -D gene also did not show, alone nor in conjunction with the -H and -I gene.

* Although our protein gels did not convey expression of neither the -I nor -D genes, our genes may still be expressed and may have enough activity to catalyze the Mg-chelatase enzyme to produce Mg-protoporphyrin IX.