Berkeley LBL/Results/ProteinGel

From 2007.igem.org

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== Discussion ==
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== '''Discussion''' ==
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Prior to running the protein gels, we expected to see protein bands for the constructs that contained the -H and -I gene(~140kDa and ~38kDa, respectively) and were also induced with IPTG .  We did not expect to see bands that coded for the -D gene (~70kDa) and for 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.  This 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; however, when 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.
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'''''Prior to running the protein gels:'''''
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''*We expected to see protein bands for the constructs that contained:''
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 +
1. -H gene (~140kDa)that were induced with IPTG
 +
 
 +
2. -I gene(~38kDa) that were induced with IPTG
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 +
''*We did not expect to see bands that coded for:''
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 +
1. -D gene (~70kDa)  
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 +
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.'''

Revision as of 03:19, 27 October 2007

Results of SDS-PAGE Gel of Soluble Proteins

Invitrogen BenchMark Protein Ladder

Plasmid Key for Protein Gels

ProteinGel1.jpg

Gel1Key.jpg

ProteinGel2.jpg

Gel2Key.jpg

ProteinGel3.jpg

Gel3Key.jpg

ProteinGel4.jpg

Gel4Key.jpg


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.

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