Cloning and Prokaryotic Expression of CDC48, and Its Expression during Somatic Embryogenesis in Dimocarpus longan Lour.

Abstract:

In order to understand the expression of CDC48 from embryo of Dimocarpus longan, a 2620 bp cDNA sequence (GenBank accession No.: EU606206) with complete ORF and a 2418 bp DNA sequence (GenBank accession No.: FJ590953) was cloned from calli of Dimocarpus longan, named DlCDC48 gene. The DlCDC48 gene is intronless, encoding 805 amino acids. Bioinformatic analysis indicated that DlCDC48 was a hydrophilic cytoplasmic protein without transmembrane domain and signal peptide, located in nucleus. The amino acid sequence had high similarity between DlCDC48 and CDC48 from other plants. Prokaryotic expression vector fused with DlCDC48 was constructed, and a protein about 89 kD was expressed after induced by IPTG. DlCDC48 expressed at all somaitc embyogenesis stages of longan by using qPCR method. The expression of DlCDC48 was the lowest at globular embryo stage, while the highest was at compact globular embryo stage. These lay a foundation for future study the CDC48 gene function during somatic embryogenesis in plants.