Molecular identification of MPaB and MPaE genes from MPA gene cluster in new strain of Penicillium brevicompactum

Abstract

Mycophenolic acid (MPA) is a fungal metabolite possessing antiviral, antifungal, antibacterial, antitumor and anti-psoriasis activities. It is being used as an immunosuppressive agent in kidney, heart and liver transplantation patients. In the presence of MPA, the proliferation of the B and T lymphocytes is inhibited. The MPaB and MPaE genes reside in a 25 kb gene cluster in the genome of Penicillium brevicompactum. In this study, the genomic DNA was extracted from P. brevicompactum grown on potato dextrose (PD) medium. To amplify the MPaB and MPaE fragments, the specific primers were designed using Gene Runner software according to P. brevicompactum IBT23078 sequence database under HQ731031.1 accession number. The amplified MPaB and MPaE genes were cloned in the PTG19-T PCR cloning vector and transformed to Escherichia coli (E. coli) top 10 competent cells. The insertion of MPaB and MPaE in the PTG19-T cloning vector was further confirmed by PCR. The MPaB and MPaE amplification produced amplicons of 1477 and 780 (nt), respectively, with the same length according to the MPaB and MpaE genes deposited in the GenBank. However, the alignment results showed some differences at nucleotide and amino acid levels, implying a new strain of P. brevicompactum.