Construction and expression of vectors encoding biologically active rodent gonadotropins

نویسندگان

  • Akihiko OHTA
  • Yuichiro TSUNODA
  • Yoshihiko TAMURA
  • Kayoko IINO
  • Naoto NISHIMURA
  • Hiroto NISHIHARA
  • Haruka TAKANASHI
  • Saishu YOSHIDA
  • Takako KATO
  • Yukio KATO
چکیده

The gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), are important hormones in vertebrate reproduction. The isolation of gonadotropins from the pituitary gland is sub-optimal, as the cross-contamination of one hormone with another is common and often results in the variation in the measured activity of LH and FSH. The production of recombinant hormones is, therefore, a viable approach to solve this problem. This study aimed to express recombinant rat, mouse, and mastomys FSH and LH in Chinese hamster ovary (CHO) cells. Their common α-subunits along with their hormone-specific β-subunits were encoded in a single mammalian expression vector. FSH from all three species was expressed, whereas expression was achieved only for the mouse LH. Immunohistochemistry for rat alpha subunit of glycoprotein hormone (αGSU) and LHβ and FSHβ subunits confirmed the production of the dimeric hormone in CHO cells. The recombinant rodent gonadotropins were confirmed to be biologically active; estradiol production was increased by recombinant FSH in granulosa cells, while recombinant LH increased testosterone production in Leydig cells.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Over Expression of Biologically Active Interferon Beta Using Synthetic Gene in E. coli

In this study, our previously reported novel synthetic gene encoding 166 residues of interferon-? was used for an efficient expression of IFN-?. The synthetic gene was cloned into pET21a expression vector and transferred into E. coli. Recombinant protein was over-expressed in the E. coli. Identity of the recombinant protein was confirmed by western blot analysis. The recombinant protein was bio...

متن کامل

Construction of Eukaryotic Expression Vectors Encoding CFP-10 and ESAT-6 Genes and Their Potential in Lymphocyte Proliferation

Background: Mycobacterium (M.) bovis is the agent of bovine tuberculosis (TB) in a range of animal species, including humans. Recent advances in immunology and the molecular biology of Mycobacterium have allowed identification of a large number of antigens with the potential for the development of a new TB vaccine. The ESAT-6 and CFP-10 proteins of M. bovis are important structural and function...

متن کامل

Transient Expression of Biologically Active Anti-rabies Virus Monoclonal Antibody in Tobacco Leaves

Background: Rabies virus is a neurotropic virus that causes fatal, but, a preventable disease in mammals. Administration of rabies immunoglobulin (RIG) is essential for the post-exposure of the prophylaxis to prevent the disease. However, replacement of polyclonal RIGs with alternative monoclonal antibodies (MAbs) that are capable of neutralizing rabies virus has been recommend...

متن کامل

Construction of New Genetic Tools as Alternatives for Protein Overexpression in Escherichia coli and Pseudomonas aeruginosa

Background: Pseudomonas protein expression in E. coli is known to be a setback due to signifi cant genetic variation and absence of several genetic elements in E. coli for regulation and activation of Pseudomonas proteins. Modifi cations in promoter/repressor system and shuttle plasmid maintenance have made the expression of stable and active Pseudomonas protein possible in bot...

متن کامل

Construction of an Eukaryotic Expression Vector Encoding Herpes Simplex Virus Type 2 Glycoprotein D and In Vitro Expression of the Desired Protein

To construct of an eukaryotic expression vector encoding herpes simplex virus type 2 (HSV-2) glycoprotein D (gD2), an Iranian isolate of HSV-2 was propagated in HeLa cell line and its DNA was extracted and used as template in polymerase chain reactions (PCR), to amplify gD2 gene. Primers were designed and the restriction enzyme sites for EcoRI and XhoI were considered at their 5′ ends respectiv...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 63  شماره 

صفحات  -

تاریخ انتشار 2017