Past Issue

Volume 9, Supplement 1, Summer 2015 (Presented at 16th Congress on Reproductive Biomedicine and 10th Royan Nursing and Midwifery Seminar) Pages: 96-96

P-127: The Effect of Beta Globin Intron on Human FSH Hormone Expression in CHO Cells


Background
Follicle stimulating hormone (FSH)- a hetrodimeric glycoprotein- is secreted by pituitary gland. This hormone stimulates growth and maturation of the follicles in females and sperms in male. Up to now, glycoprotein hormones such as FSH have produced in different cell lines. Among of the mammalian expression systems, the Chinese hamster ovary cells (CHO) have taken into consideration owing to high level of expression and post-translational modifications on proteins. Nowadays, recombinant FSH is produced in CHO cells. One of the pathways of gene expression optimization is use of introns. Introns are non-coding sequences that is located between coding sequences in the majority of eukaryotic genes. Human beta globin gene is located on chromosome subband 11p15.5. This gene is composed of 3 exons and 2 introns. The length of intron is 130 and 850 base pairs, respectively. The aim of this study was to investigate the effects of intron I beta globin gene on FSH expression.
Materials and methods
At first, the gene construct was made by PCR and SoeingPCR. Then, this structure was cloned into the pTZ57R/T vector and pVitro2-neo-mcs expression vector.
Results
In continuous, recombinant clones were confirmed by colony PCR and sequencing techniques.
Conclusion
In the future, this gene construct will be transformed into CHO cells and exmined the expression level of recombinant protein by SDS-page and Western blotting methods.