Abstract: Glycolate oxidase 1(GOX1) is one of the key enzymes in the photorespiration process of Arabidopsis thaliana, which can oxidize glycolic acid to glyoxylic acid and hydrogen peroxide. In this paper, wild-type Arabidopsis thaliana Columbia(Col-0) was used as the research material. The 1 104 bp CDS sequence of GOX1 gene was obtained by reverse transcription-polymerase chain reaction (RT-PCR) technology, its gene function was analyzed preliminarily using bioinformatics technology, PET28a(+)-GOX1 prokaryotic expression vector with His tag was transformed into E. coli competent BL21, and SDS-PAGE electrophoresis was used to detect the protein product after induced expression. The GOX1 gene encodes 367 amino acids, the relative molecular mass is 40 341.48, and the theoretical isoelectric point is 9.16. It is a hydrophilic protein and is located in peroxisomes. SDS-PAGE detected the band of interest at 40 kDa, which was consistent with the theoretical prediction. Through the analysis of GOX1 gene expression, it has laid the foundation for further exploration of protein function and regulation mechanism.
Keywords: glycolate oxidase 1(GOX1); gene cloning; bioinformatics analysis; prokaryotic expression
合肥工业大学学报自科版