Molecular cloning and blast by gene bank of alpha and beta subunit in LH hormone into mammalian shuttle vectorPEGFP-N1 using IRES

Background and Objective: The glycoprotein hormone secreted from anterior pituitary gland are heterodimeric, non- covalently consisting of a common α subunit and a hormone-specific β subunit. Due to the lack of adequate access to hormones and the possibility of contamination, this study attempted to clone two subunits of LH hormone by using IRES sequence in eukaryotic expression vector pEGFP-NI for clinical purposes.

Materials and Methods: To clone the LH hormone, we designed the subunits alpha and beta sequences, then synthesized the plasmid vector pGEM-BI. The designed sequence was subcloned in the SmaI site of expression plasmid pEGFP-N1. The resulting plasmid was confirmed by restriction enzyme digestion and PCR.

Results: PCR and restriction enzyme analysis showed that the plasmid pEGFP-Nl-α-IRES-β contains the correct sequence of the target gene sequences in the gene bank.

Conclusion: Since the plasmid has a proper structure, it can be used to transfer into eukaryotic systems and is also appropriate for the evaluation of expression.