Identification and characterization of a heat-inducible Hsp70 gene from Sorghum bicolor which confers tolerance to thermal stress

摘要

This study describes the first detailed molecular characterization of the heat shock protein 70 (Hsp70) gene from Sorghum bicolor, MN1618 designated as SbHsp70-1. The full-length cDNA of SbHsp70-1 consists of 2524?bp with a 1950 bp open reading frame, which encodes a protein of 649 amino acids. SbHsp70-1 is a cytoplasmic protein with high homology to other plant Hsp70s, especially grain crops. Recombinant SbHsp70-1 was able to bind and hydrolyse ATP in a dose-dependent manner, suggesting that SbHsp70-1 functions as an ATPase. Immunoblot assays showed that the expression of SbHsp70-1 is induced at temperatures of 37, 45, and 4?°C but reduced at 42?°C. In addition, the SbHsp70-1 mRNA transcript is constitutively expressed in both leaves and stem but is significantly increased upon heat shock at 42?°C. Upon cold shock at 4?°C, SbHsp70-1 mRNA transcript level increased in the leaf, but no significant change was observed in the stem. In addition, expression of the pET28a-SbHsp70-1 construct in Escherichia coli cells under heat stress resulted in their survival even at higher temperature (65?°C). Our results suggest that SbHsp70-1 is a heat-inducible protein that confer thermal tolerance to bacterial cells and can be claimed as a promising target to study stress tolerance in crops.