Isolation and characterization of abiotic stress related gene (cspA) from Enterobacter cloacae
Student name: Ms Tilky Singh
Guide: Dr Anandita Singh
Year of completion: 2014
Host Organisation: National Research Centre on Plant Biotechnology, New Delhi
Supervisor (Host Organisation): Dr Kanika
Abstract: Abiotic stresses including cold stress are serious threats to the sustainability of crop yields
accounting for more crop productivity losses in agriculture. The genes conferring tolerance to
abiotic stress can be isolated, cloned and introduced into important crop plants. CSPs (cold shock
proteins) in microbes are induced during cold shock and these CSPs are also associated with the
cold stress tolerance in plants. Cold shock inducible genes such as cspA and cspB functions as RNA
chaperones to prevent the formation of secondary structures in RNA molecules at low
temperatures. Cold shock domain proteins (CSDPs) have been cited in different plant species which
perform similar functions to CSPs in prokaryotes during cold acclimation. The present study was
conducted to clone cspA gene from microbial isolate (Enterobacter cloacae) and transform it into a
model plant i.e. Arabidopsis thaliana, ecotype Columbia. The amplified cspA gene (213 bp) was
successfully cloned in pDrive cloning vector. Sequence analysis of cspA gene isolated from
Enterobacter cloace shows its maximum identity with the cspA genes of other bacteria like
Pseudomonas, Salmonella typhimurium, Escherichia coli, etc., cspA was sub-cloned into binary
vector in sense direction downstream to 35S promoter and transformed into Agrobacterium
tumefaciens strain (GV3101). Transgenic Arabidopsis plants were developed with the help of
Agrobacterium mediated in-planta transformation. Further, cspA gene was also sub cloned in
pET29A expression vector for its protein expression studies. This novel gene can be an important
component of possible utilization in abiotic stress management for crop plants.
Keywords: Abiotic stress, Cold shock proteins, cspA, RNA chaperons,
Enterobacter cloacae
