Cloning And Its Prokaryotic Expression Of Heat Shock Protein Gene From Capparis Spinosa

Heat shock proteins are the most important adversity induced proteins in plant. They are speedy synthesized and protected cell avoid damage when plants suffering adversity especislly temperature stress. Depend on their molecular chaperone function they participated in various cellular processes under normal and stress conditions. They are also playing key role in the processes of plants defend environment stress such as heat, cold and oxidation. Accordingly, the heat shock genes are fine genes resources which have important practical valuable in breeding work. As the special plants which were found in aridity zone, Capparis spinosa has its strong drought adaptability with strong resistance of aridity, high or low temperature, wind corrode and barren. The base environment they lived always associate with primitive desert soil in aridity background. These characteristic determined that it is the excellent genes donor to breeding research. Therefore, it has important practical significance to developing and using these fine gene resources of Capparis spinosa.RNA isolation is the most important step in genetic engineering. The success or failure of gene clone has key relations with quality of RNA. An effective improved SDS method of isolation of total RNA from leaves of wild Capparis spinosa was developed. The RNA extracted by this method was with low cost, high yield and fast.By the methods of RT-PCR and rapid amplification of cDNA ends (RACE) we succeed isolated a HSP70 gene (CsHSP70) and a sHSP gene (CsSHSP) from lamina of Capparis spinosa and logined to Genebank with the accession numbers EU574936 and EU574935. The CsHSP70 gene contains 2202 nucleotides with a 1950 nucleotides length open reading frame (ORF) coding 649 amino acid residues with the predicted molecular weight of 71.0446 kD. This gene belongs to HSP70 gene family. The CsSHSP gene contains 917 nucleotides with a 678 nucleotides length open reading frame (ORF) coding 225 amino acid residues with the predicted molecular weight of 25.119 kD. This gene belongs to chloroplast sHSP gene family.Recombinant CsHSP70 and CsSHSP were overexpressed in Escherichia coli. By the temperature treatment experiment we found that these recombinants can improved the capacity of Escherichia coli to against high (50℃) and low (4℃) temperature.