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Abstract
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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.
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