1. department of biology. faculty of science. urmia university. urmia.iran 2. department of biology. faculty of science. urmia university. urmia.iran
Abstract
Introduction
: mycophenolic acid (mpa) is a fungal metabolite possessing antiviral, antifungal, antibacterial, antitumor and antipsoriasis activities. it is being used as an immunosuppressive agent in kidney, heart, and liver transplantation patients. in the presence of mpa, proliferation of b and t lymphocytes is inhibited. the mpae gene resides in a 25kb gene cluster in the genome of penicillium brevicompactum.
Methods
Genomic dna was extracted from penicillium brevicompactum grown on pd medium. in order to amplify the mpae fragment, gene specific primers were designed using gene runner software according to penicillium brevicompactum ibt23078 sequence database under hq731031.1 accession number. the amplified mpae gene was cloned in to ptg19-t pcr cloning vector and transformed into ecoli top 10 competent cells.
Results
The insertion of mpae into the ptg19-t cloning vector was further confirmed by pcr. after the selection of positive screen colonies using colony-pcr, the sequencing of the recombinant plasmids was performed. the obtained sequence was analyzed using blast and clustalw software program. this sequence was compared and aligned with other mpae sequences in different species. results showed that this sequence had differences with other fungal species. according to the results of the current study a new sequence from strain mpae has been published for the first time.
Conclusion
The mpae amplification produced an amplicon (~780 nt) larger than mpae gene reported in database, implying a new strain of penicillium brevicompactum.
