Cloning of the phosphoenolpyruvate carboxylase gene (PEPC) from Isoetes shangrilaensis X. Liu and construction of its expression vector

Using Chinese Isoetes shangrilaensis X. Liu, the phosphoenolpyruvate carboxylase gene (IsPEPC) sequence information was screened by transcriptome sequencing analysis, and the IsPEPC gene encoding the phosphoenolpyruvate carboxylase (PEPCase) was cloned from cDNA based on the gene sequence. The IsPEPC gene was inserted into the pCAMBIA-2300-N-eGFP and pMD plasmid vectors, after which the recombinant vectors were transferred into wild-type Arabidopsis thaliana (L.) Heynh. by agrobacterium-mediated inflorescence infiltration. Results showed that the protein-coding sequence of the IsPEPC gene was 2928 bp in length and encoded 975 amino acids. Homologous searching showed that the protein was related to the PEPC protein sequence of the source species Selaginella moellendorffii Hieron., with 79.8% sequence homology. Transgenic T₁ A. thaliana was screened for resistance and positively identified at the gDNA level. We initially identified 26 pC2300-N-eGFP-IsPEPC transgenic lines and 32 pMD-IsPEPC transgenic lines.