Part:BBa_K1526005

LacRS + Gsh1*

Lac Repressor System + Gsh1

Original Gene: gsh1/gshA
Organism: Saccharomyces cerevisiae
Protein: Glutamate-Cysteine Ligase (previously known as gamma-glutamylcysteine synthetase)
Function: Gamma glutamylcysteine synthetase catalyzes the first step in glutathione (GSH) biosynthesis;
L-glutamate + L-cysteine + ATP -> gamma-glutamyl cysteine + ADP + Pi
Expression is induced by oxidants, cadmium, and mercury. Protein abundance increases in response to DNA replication stress.
Aim: We can use the overproduced cysteine to make gamma-glutamyl cysteine, which is the monomer that forms phytochelatins (n=10-20). In order to do this, we need glutamate-cysteine ligase to catalyse the reaction. We can over-express GSH1* using the pYodA promoter.
Mutant strains in E. coli: Information about mutant strains for Gsh1 from the Keio collection can be found at [http://www.york.ac.uk/res/thomas/Gene.cfm?recordID=EB0413&CFID=9377833&CFTOKEN=d775499c2963924f-23DC4F02-CF19-BFB5-F290A2DBAAB49113&jsessionid=b63022c18b939163d6b5485d476425935481TR EchoBase website] hosted by the University of York

Literature:

1. http://biocyc.org/YEAST/NEW-IMAGE?type=GENE-IN-MAP-IN-PWY&object=YJL101C

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Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1207
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1800
  
• 1000
  COMPATIBLE WITH RFC[1000]