[Objective] The aim was to study the expression of Arabidopsis gene A/2G34450 in Pichia pastoris and to obtain recombinant Arabidopsis HMGB protein. [Method] The At2G34450 gene was cloned into yeast expression vector pPIC9K containing AOXl promoter and the sequences of secreting α-signal peptides. Recombinant plasmid was linearized by Sal l and transformed into P. pastoris GSl15 competent cells by electroporation. Positive integrated clones were screened out, and the At2G34450 protein was expressed under the induction of methanol. [Result] The At2G34450 protein was expressed in yeast medium through methanol induction. SDS-PAGE results showed that recombination product was At2G34450 protein. [Conclusion] At2G34450 protein was successfully expressed in the P. pastoris system for the first time, which paves a direct path to further research on the functions of HMGB family members.
[Objective] The aim was to better research the function and action mode of High Mobility Group B （HMGB） proteins in higher plants. [Method] At2G33450,At5G23405 and At5G23420 genes of HMGB protein family in Arabidopsis thaliana were cloned by the use of RT-PCR method,and the expression of these three proteins in E.coli and Arabidopsis thaliana were detected by using SDS-PAGE,Northern blot and subcellular localization methods. [Result] The results showed that the molecular weights of the three proteins were 17.5,17.0 and 27.0 kD respectively,and the expression levels of the proteins in Arabidopsis thaliana were At5G23420At5G23405At2G33450. In addition,all the three proteins were located in nucleus. [Conclusion] The study will provide a basis for the further research on the biological function of HMGB proteins in higher plants.
