For amplification PCR was performed by using specific primers for glutathione synthetase gene.
Sequence of Glutathione Synthetase gene in the reference strain Synechocystis sp.
The query asking glutathione synthetase gene from CyanoBase (The Genome Database for Cyanobacteria) for the reference strain Synechocystis sp.
Term: 1052121 Length(aa):320Direction: direct Gene Products less than less than Gene Name greater than greater than : glutathione synthetase less than less than gshB greater than greater than The gene sequence was Synechocystis (Chr) Synechocystis PCC 6803 (Chr) 1051159-1052121"
Features in this part of subject sequence: Solanesyl diphosphate synthase Glutathione synthetase
Amplification of Glutaredoxin encouraged for the isolation, amplification and sequencing of glutathione synthetase gene.
Glutathione synthetase (gshB) gene analog of Synechocystis PCC 6803 was sequenced in Synechocystis AHZ-HB-MK" and Synechocystis AHZ-HB-P2A".
Almost in all cyanobacterial strains a polypeptide of 35 KDa was found, which may be the product of gshB gene responsible for the synthesis of glutathione synthetase.
The failure of gene fragment amplification in cyanobacterial stains analogue to glutathione synthetase (gshB), except Synechocystis AHZ- HB-MK" and Synechocystis AHZ-HB-P2A" strains, may have one explanation that the primers designed for glutathione synthetase gene probably was not form conserved region of this gene, in other cyanobacteria (Jeff Elhai, personal communication), so Synechocystis strains gave amplification, while other cyanobacterial strains did not give amplification for it.
Two genes glutathione synthetase and glutaredoxin 3 are involved in glutathione synthesis and reduction of oxidized form of glutathione (GSSG in to GSH) to reduced form for its further use to overcome oxidative stress by binding with heavy metals .
Strain Synechocystis AHZ- HB-MK" is not only different from Synechocystis AHZ-HB- P2A" on the bases of glutathione synthetase gene homology which is greater in case of Synechocystis AHZ-HB-MK", but also from the other aspects.