Past Issue

Volume 9, Supplement 1, Summer 2015 (Presented at 16th Congress on Reproductive Biomedicine and 10th Royan Nursing and Midwifery Seminar) Pages: 94-94

P-122: The Effect of Beta Globin Intron on Human LH Hormone Expression in CHO Cells


Background
Luteinizing hormone (LH) is a heterodimeric glycoprotein composed of alpha and beta subunits.This hormone is secreted from the pituitary gland. LH, in women triggers Menstrual cycle and ovulation. In men, LH stimulates production of testosterone, which plays a specialized role in sperm production. Up to day, LH hormone have produced in different ways such as codon optimization and also in various cell lines. Between mammalian expression systems, Chinese hamster ovary (CHO) cells, due to rapid proliferation and post-translational modifications, are more common than other hosts. One way to increase gene expression is use of introns. Beta-globin intron is composed of 3 exons and 2 introns. Human beta globin gene is located on chromosome subband 11p15.5. In this study, the beta-globin gene intron-I was used to enhance the expression of LH hormone. The purpose of this study, was to investigate the effects of intron I beta globin gene on LH expression.
Materials and methods
The gene construct was made by PCR and Soeing-PCR techniques. Then, this structure was cloned into pVitro2-neo-mcs expression vector.
Results
In continuous, recombinant colonies were approved by colony PCR, digestion and sequencing.
Conclusion
In the future, gene construct will be transferred into CHO cells by electroporation technique and exmined the expression level of recombinant protein by SDS-page, Western blotting and ELISA methods.