Paper Information

Journal:   INTERNATIONAL JOURNAL OF FERTILITY AND STERILITY   SUMMER 2012 , Volume 6 , Number SUPPLEMENT 1; Page(s) 121 To 121.
 
Paper: 

EXPRESSION OF BOVINE FOLLICLE STIMULATING HORMONE SUBUNITS IN PICHIA PASTORIS

 
 
Author(s):  EBRAHIMI S.*, GOURABI H., AMIRI YEKTA A., FATEMI N., ESKANDARI NASAB M.P., BAHRAMINEJAD E., AHANJAN M., MIRSHAHVALADI SH., NEKOOKAR A.B., SANATI M.H.
 
* DEPARTMENT OF GENETICS, REPRODUCTIVE BIOMEDICINE RESEARCH CENTER, ROYAN INSTITUTE FOR REPRODUCTIVE BIOMEDICINE, ACECR, TEHRAN, IRAN
 
Abstract: 
Background: Bovine follicle stimulating hormone (bFSH) belongs to the family of glycoprotein hormones that are released from the pituitary gland or the placenta. This hormone is a heterodimer consisting of common a- and specificb-subunit. Bovine FSH is used for reproductive technology such as superovulation in farm animals and polycystic ovary syndrome treatment. Also this hormone is produced for clinical, veterinary purposes and infertility treatment.
Recombinant DNA technology provides a useful tool for production of FSH free from hormonal, viral or prions contaminants. The objective of our study was the production of bFSH hormone in Pichia pastoris expression system.
Materials and Methods: For this purpose, two ORF strands containing
a and b subunits were separately cloned into pTZ57R/T vector and then were subcloned into pPIC9 expression vector and confirmed by PCR and sequencing.
The two recombinant plasmids were linearized using the SalI and SacI restriction enzymes in order to generation of His+Mut+GS115. The products were co-transfected into the competent yeast cells by electroporation and examined by PCR.
Results: Expression of recombinant bovine FSH in both yeast cells supernatant and pellet was confirmed by SDS-PAGE and Western blotting techniques.
Conclusion: Production of recombinant protein in eukaryotic expression systems such as pichia pastoris is a reliable method for producing of therapeutic bovine FSH.
Also the detection of the recombinant produced protein by specific antibody confirmed a correct post translated modified structure.
 
Keyword(s): BOVINE FOLLICLE STIMULATING HORMONE, EXPRESSION, PICHIA PASTORIS
 
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