Abstract:

PGIP is a cell wall binding protein specific binding and inhibiting activity of fungal endo-PG activity and occupies a very important position in plant molecular resistance breeding. Cloning and sequence analysis of PGIP gene in Brassica oleracea L. will lay the foundation for breeding resistant disease varieties through the transgenic technology. Therefore, in the paper, through the homologous sequence cloning method, a new PGIP gene, designed as BoPGIP, were cloned from Brassica oleracea L. by PCR amplification, with two pairs of gene specific primers designed according to the full-length sequences of known BoPGIP1 gene (Accession: JF325875) from Brassica oleracea var. italica. The DNA full-length of BoPGIP was 1065 bp and consisted of one intron and two exons. The full length of the exons was 975 bp and encoded a protein of 342 amino acids with a predicted molecular mass of 43.8 kD and a 6.256 Isolectric Point. Bioinformatics analysis revealed that the BoPGIP gene belonged to the PGIP gene family and have typical LRR domains; moreover, it had four potential N-glycosylation sites (65-68, 106-109, 130-133, 254-257), one protein kinase C phosphorylation site (189-191), five casein kinase II phosphorylation sites (73-76, 78-81, 151-154, 182-185, 304-307) and four N-myristoylation site (157-162, 188-193, 236-241, 273-278); signal peptide analysis showed BoPGIP might encode a membrane protein with a signal peptide at the N-terminus. The predicted secondary structure composition for the protein had 31.79% helix, 13.89% sheet, and 54.32% loop. These results will lay the groundwork for further studying the function of the BoPGIP gene. Cloning and sequence analysis of BoPGIP gene revealed that it belonged to PGIP gene family and will provide a basis for further verify its function.