Construction of Geobacillus thermoglucosidasius cDNA library and analysis of genes expressed in response to heat stress

• DOI: 10.1007/s11033-013-3011-7
• Publication Date: 2014-01-07T17:48:56Z
• Authors: S. Tripathy; N. K. Maiti
• Keywords: Expressed Sequence Tags; 0301 basic medicine; 03 medical and health sciences; Hot Temperature; Bacterial Proteins; Geobacillus; Gene Expression Regulation, Bacterial; RNA, Messenger; Sequence Analysis, DNA; Heat-Shock Response; Gene Library

Thermophiles exhibit various kinds of molecular mechanisms to survive in extreme environment, but their behavioral responses to long duration stress is poorly understood until date. In the present study, we have prospected for the genes differentially expressed in response to long duration heat stress in thermophilic bacteria. A cDNA library was constructed from Geobacillus thermoglucosidasius grown with a temperature upshift of 10 °C from optimum growth temperature of 45 °C for 16 h. A total of 451 clones from the library were sequenced with accurate base calling that generated 257 high quality sequences with an average read length of 350 bp. We queried our collection of single pass sequences against the NCBI non-redundant database using the BLASTX algorithm and obtained sequences that showed significant similarity (>60%) with heat shock proteins, metabolic proteins and hypothetical proteins. The expressed sequence tags (ESTs) expressed in response to heat stress were annotated that further commuted a strong interaction network among one another. The ESTs based on the best hits were validated by RT-PCR. Di- and tri-nucleotide repeat motifs were also found to be associated with 17 genes involved in heat shock response, metabolism, transport and transcriptional regulation. The present results provide the novel identification of the putative genes responsible for imparting tolerance to bacteria under heat stress and unveil their role for survival of life in environmental extremes.