The Expression And Cloning Research Of Gene Tps1 In Yeast

Under the condition of nature, yeasts are exposed to a highly variable environment with respect to the availability and quality of nutrients, temperature, pH, radiation, access to oxygen, and water activity. But yeast has one of the precise system to adapt to the variable environment after long evolution. Trehalose, as an important stress-resistance factor, play a key role in the yeast to resistant to the various stress. As a key process, UDP-glucose is converted first to trehalose-6-phosphate by trehalose-6-phosphate synthase (TPS1) and then to trehalose. As a result, it is remarkable to research the expression of gene tpsl.In this paper, three yeast stock, which have different growth properties, are used as material. After the cloning of gene tpsl, sequencing revealed that there are eight mutation site of WFB tpsl, and accordingly, FHS is nine and YY is thirteen.On the other hand, the protein sequence of TPS1 of FHS and WFB haven't any difference compare to one protein sequence of TPS1 reported by GeneBank. But the TPS1 sequence of YY has three mutations sites. The locus of three mutation site are 67, 69 and 388. Based on the analysis of conserve sequence of protein, finding reveals that the mutation is remarkable.After heat shock, the results of Dot Bloting reveal that tps1 mRNA of three yeast have different expression level. Under the condition of 37°C , tps1 mRNA of FHS and WFB have a high abundance, but tpsl mRNA of YY is low. When subject to 42°C, the expression of tps1 mRNA of FHS and WFB reach their maximum. Correspondingly, YY reach its maximum at 48 °C.When subject to the solution of NaCL, the abundance of tpsl mRNA of three yeast have no remarkable change. This result prove trehalose have less relative to theresistance of yeast to the stress of high external osmolarity...