Cloning expression and analysis of phytochelatin synthase (pcs) gene from Anabaena sp. PCC 7120 offering multiple stress tolerance in Escherichia coli.
Phytochelatin synthase (PCS) is involved in the synthesis of phytochelatins (PCs), plays role in heavy metal detoxification. The present study describes for first time the functional expression and characterization of pcs gene of Anabaena sp. PCC 7120 in Escherichia coli in terms of offering protection against heat, salt, carbofuron (pesticide), cadmium, copper, and UV-B stress. The involvement of pcs gene in tolerance to above abiotic stresses was investigated by cloning of pcs gene in expression vector pGEX-5X-2 and its transformation in E. coli BL21 (DE3). The E. coli cells transformed with pGEX-5X-pcs showed better growth than control cells (pGEX-5X-2) under temperature (47 degrees C), NaCl (6% w/v), carbofuron (0.025 mg ml(-1)), CdCl2 (4 mM), CuCl2 (1mM), and UV-B (10 min) exposure. The enhanced expression of pcs gene revealed by RT-PCR analysis under above stresses at different time intervals further advocates its role in tolerance against above abiotic stresses.
Chaurasia
Neha
N
Molecular Biology Section, Laboratory of Algal Biology, Center of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005, India.
Mishra
Yogesh
Y
Rai
Lal Chand
LC
D019881:Aminoacyltransferases / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D017033:Anabaena / Q000201:enzymology* / Q000235:genetics; D019256:Cadmium Chloride / Q000494:pharmacology; D003001:Cloning, Molecular; D003300:Copper / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000528:radiation effects; D017343:Genes, Plant / Q000502:physiology*; D006358:Hot Temperature; D054811:Phytochelatins / Q000096:biosynthesis; D012965:Sodium Chloride / Q000494:pharmacology; D014466:Ultraviolet Rays
D012965:Sodium Chloride; D003300:Copper; D054811:Phytochelatins; D019881:Aminoacyltransferases; C093784:glutathione gamma-glutamylcysteinyltransferase; D019256:Cadmium Chloride