The cloning of Fusarium polygalacturonase gene fragments using degenerate primers designed by CODEHOP

With the help of CODEHOP,degenerate primers were designed for cloning the fungal polygalacturonase gene fragment.The proper primer was screened from many pairs of primers.TD-PCR and the popular PCR reaction were compared to determine the good reaction system.The result showed that a 700 bp PCR product with a length close to the target gene fragment was obtained,the TD-PCR with the same reaction system has higher specificity than the popular PCR reaction system,and the amino acid fragments encoded by the products of the cloned DNA had high similarity with those of polygalacturonase amino acid sequence from GenBank.The cloned sequence should be polygalacturonase gene fragment from the tested Fusarium.This study indicated that the degenerate primers designed by the CODEHOP software are practicable and high in positive rates.Cloning of pectinase gene fragment of the tested Fusarium would afford a scientific warrant and possibility for the research of pathogenic mechanism of Fusarium.