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Abstract Genomic DNA of pullulanase (EC 3.2.1.41), glycosyl hydrolase family 57 from Aureobasidium pullulans, was determined using polymerase chain reaction, according to Baser program, gave assembly for 16 sequenced samples from 22 samples depending on identity percent, overlap and assembly scores, different amplified fragments were sequenced as 2604 bp. and G+C content is 51.8 % with molecular mass 80 KDa. Pullulanase gene determines 8 exons, splice 1 from (312 – 246), splice 2 from (516 - 685), splice 3 from (1064 - 1161), splice 4 from (1929 - 1218), splice 5 from (1298 - 1349), splice 6 from (1932 - 2102), splice 7 from (2286 - 2173) and splice 8 from (2513 - 2422).This is the first time to sequenced the pullulanase from Aureobasidium pullulans. Nine restriction enzymes (RE) were used to predicts five open reading frame (ORF), first restriction enzyme used #BssKI gave a 147 amino acids, the second RE were AluI and Tsp509I gave 142 amino acids, third RE were *BceAI and FatI gave 120 amino acids, fourth RE were *NciI and DdeI gave 116 amino acids and fifth RE were *BtgZI and MseI gave 100 amino acids. Translation of nucleotide sequence resulted 867 amino acid using frame 2. Pullulanase nucleotide collection from Aureobasidium pullulans was blasted which showed high similarity with S. oleracea L. 88% and Ricinus communis 81% using NCBI significant alignment. EMBL- EBI tools, ClustalW2 scores for pullulanase nucleotides from Aureobasidium pullulans was 54% in comparison to both S. oleracea and Zea mays. |