ماندانا زارعی

Making super organism has always been one of the human goals. A method to achieve this aim is biotechnology. Biotechnology through making organisms with more growth enhancement, more tolerant to acute conditions and Pathogens, provides a way to reach this goal. Expression vectors are one of the crucial tools for making transgenic organisms and the most important elements of this tool is promotor. Promotor determines when and how RNA polymerase binds to DNA. In aquaculture, most attention had been toward viruses and mammalian promotors but by passing time and more studies their defect were understood and researchers considered promotors with aquatic origin. Beta-actin promotor is one of the promising choices in this part. This promotor causes high expression of beta-actin in organism’s body and its expression is permanently and non-inducible. In this research, the isolation of penaeus semisulcatus bet-actin promotor was investigated using semiAFLP, RS-PCR, mRS-PCR and Inverse PCR methods. Genomic DNA was extracted using methods such as CTAB, TES and Dellaporta and also commercial kits were used. This research showed RS-PCR and mRSPCR are not promising. A 350 bps segment was achieved using IPCR but after analyzing was specified, it is downstream of promotor. With alteration in semiAFLP, a longer segment was gained that had 500 bps. In this method, at first, genomic DNA was digested with B, H and S enzymes. Then the adaptors attached. In first round PCR a sharp band was seen on the agaros gel. This result was confirmed by nested PCR. Achieved segments were sequenced and their homologies were compared using Blast software. Blast results showed two segments have homology with L. vnnamei beta-actin promotor.