关键词: 拟南芥, PRF1, 瞬时表达, 原生质体, GFP

Abstract: Here we study the subcellular localization of profilin-1（PRF1）using two transient expression methods and compare the advantages and disadvantages of two transient expression methods in the subcellular localization. Using the leaves of Arabidopsis thaliana as material,the total RNA from the leaves was extracted,and PRF1 gene was cloned using specific primers by RT-PCR and then ligated to pCAMBIA1300-GFP vectors,finally one plant expression vector pCAMBIA1300- GFP-PRF1 was constructed successfully. Two transient expression systems,agroinfiltration of tobacco leaves,and PEG transformation of protoplasts isolated from the leaves of Arabidopsis thaliana were adopted. The expression of the fusion proteins was observed by a confocal laser scanning microscopy. The results showed that the green fluorescence of the fusion protein was observed when the PRF1 gene was introduced into the protoplasts of A. thaliana and tobacco epidermal cells,the product of PRF1 gene and GFP fusion protein was mainly localized in the cytoplasm of tobacco epidermal cells,and there was also an expression in the nuclear cell organelle. PRF1 was localized in the nucleus and cytoplasm of Arabidopsis thaliana. The localization of PRF1 protein was different in two different transient expression systems,which may be related to the characteristics of the plants that are homologous or heterologous expression.

Key words: Arabidopsis thaliana, PRF1, transient expression, protoplasts, GFP