Berkeley LBL/Results/ProteinGel

From 2007.igem.org

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('''Discussion''')
('''Discussion''')
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''*We did not expect to see bands that coded for:''
''*We did not expect to see bands that coded for:''
   
   
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1. -D gene (~70kDa)  
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* 1. -D gene (~70kDa)  
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2. - All constructs that were not induced with IPTG
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* 2. - All constructs that were not induced with IPTG
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'''''After running the protein gels:'''''
*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.   
*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.   

Revision as of 03:21, 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

After running the protein gels:

  • 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.

Conclusion:

* 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.