Part:BBa_K5327022

Glutathione S-transferase F9

Function:

In vitro, possesses glutathione S-transferase activity toward 1-chloro-2,4-dinitrobenzene (CDNB) and benzyl isothiocyanate (BITC), and glutathione peroxidase activity toward cumene hydroperoxide and linoleic acid-13-hydroperoxide. May be involved in the conjugation of reduced glutathione to a wide number of exogenous and endogenous hydrophobic electrophiles and have a detoxification role against certain herbicides.

Usage and Biology

Genome localization:Chromosome 2 - NC_003071.7

Expression diagram:

Fig 1. The expression diagram of glutathione S-transferase F9

Corresponding enzyme structure:

Fig 2. The corresponding enzyme structure of glutathione S-transferase F9

The PCR result:

Fig 3. The PCR result of glutathione S-transferase F9

Subcellular localization:

Located in the cytoplasm of cells

Fig 4. The subcellular localization of glutathione S-transferase F9

Dynamics data:

Table 1. The dynamics data of glutathione S-transferase F9

Design Notes

The Glutathione S-transferase F9 (GSTF9) gene was designed based on the coding sequence (CDS) from Arabidopsis thaliana and codon-optimized for Saccharomyces cerevisiae (S288C) to ensure efficient expression in yeast. GSTF9 exhibits glutathione S-transferase activity toward 1-chloro-2,4-dinitrobenzene (CDNB) and benzyl isothiocyanate (BITC), as well as glutathione peroxidase activity toward cumene hydroperoxide and linoleic acid-13-hydroperoxide in vitro. It is likely involved in the conjugation of reduced glutathione to a variety of hydrophobic electrophiles and plays a detoxification role against certain herbicides. In this experiment, the GSTF9 gene is expressed in yeast using the GPD promoter (GPDp BBa_K517001) and ADH1 terminator (ADH1t BBa_K2637012) to ensure high expression levels and mRNA stability. The optimized gene is inserted into a vector and integrated into S. cerevisiae S288C via homologous recombination for expression verification. This design aims to enhance GSTF9 expression in yeast, thereby improving its detoxification capacity and optimizing yeast as a metabolic engineering platform.

Plasmid

Fig 1. The plasmid expression of glutathione S-transferase F9

Source

Arabidopsis thaliana

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 82