Sub-cloning of alpha subunit of anterior pituitary glycoprotein hormone into a mammalian expression vector

How to Cite:Mohammad Reza AkbariDidgahireza nasrAliAkbar ShabaniMojtaba GhadamYariSub-cloning of alpha subunit of anterior pituitary glycoprotein hormone into a mammalian expression vector.koomesh.13(2):e152501.

Abstract

Introduction: Anterior pituitary glycoprotein hormones include thyroid stimulating hormone, lutinizing hormone, follicule stimulating hormone, and gonadotropine hormone. Each of them contains alpha and beta subunits. The alpha subunit gene is the same in all of these hormones and contains 4 exones and 3 intrones. The beta subunit is responsible for specific function of each hormone. The aim of this study was to clone alpha chain cDNA of glycoprotein hormones in a proper vector for eukaryotic system. Material and Methods : To clone cDNA, alpha subunit of glycoprotein hormones was amplified by using one pair primers and T.vector as template and cloned in Not I and Bam HI sites of pcDNA3.1 plasmid. The recombinant plasmid transformed to E.coli Top10F΄ cell and colonies that contain plasmid were selected by Colony PCR. The accuracy of extracted plasmid of these clones was approved by enzyme digestion and sequencing. Results : Enzyme analysis showed that pcDNA3.1-F351 a had correct structure and sequencing confirmed by 100% homology of the gene with reported alpha gene in Gene Bank. Conclusion : Because of its proper structure, this plasmid is able to transform to Eukaryotic system and translation.

Keywords

Follicle stimulating hormone, Pituitary hormones, Glycoprotein hormones, Alpha subunit, Mammals

هورمون محرک فولیکولی، هورمون های غده هیپوفیز، هورمون های گلیکوپروتئینی، زیر واحد آلفا، پستانداران

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