Abstract
ABSTRACT Insulin-like growth factor-1 (IGF-1) is regarded as a crucial clinically significant therapeutic agent against several pathological conditions. Recently, recombinant DNA (rDNA) technology has enabled the production of many drugs of rDNA-origin including IGF-1. Securing a readily available supply of IGF-1 is invaluable to clinical research and biotechnological domains. In this work, the cloning of a full-length bovine IGF-1 cDNA and the successful expression of its cognate recombinant IGF-1 protein is reported. Single-strand cDNA was prepared from liver tissues, through the specific reverse transcription (RT) of IGF-1 mRNA. Subsequently, a PCR amplicon of ~543bp was successfully amplified. Recombinant pTARGET™ vector harboring IGF-1 insert was successfully cloned into competent E. coli JM109 cells. SDS-PAGE analysis revealed that the recombinant IGF-1 has been expressed at the expected size of 7.6kDa. The outcome provides a robust basis for transecting the recombinant pTARGETTM vector, harboring the IGF-1 cDNA insert, into mammalian cells. Optimal initial glucose concentration was found to be 10g/l with corresponding protein concentration of 6.2g/l. The proliferative biological activity crude recombinant IGF-1 protein was verified on HeLa cell lines. This is envisaged to facilitate large-scale production of recombinant IGF-1 protein, thereby enabling thorough investigation of its clinical and pharmaceutical effects.
Highlights
Insulin-like growth factors (IGFs) constitute a family of structurally-associated polypeptides controlling several developmental aspects in mammalians
Insulin-like growth factor-1 (IGF-1) that essentially functions as an endocrine hormone which is primarily secreted by hepatocytes and is subsequently transported to other organs (Laron, 2001)
We targeted liver tissues for the cloning of IGF-1 cDNA due to the expected high abundance of its mRNA. This is the first report on the cloning and protein expression of IGF-1 from Egyptian bovine population
Summary
Insulin-like growth factors (IGFs) constitute a family of structurally-associated polypeptides controlling several developmental aspects in mammalians. Expression of IGFs gene family and their cognate mRNA transcripts have been observed in tissues and embryos of diverse species (Naicy et al, 2017). To this end, mounting evidence suggest that the IGFs play a crucial part during early growth and development both in vitro and in vivo (Jansen et al, 1983). In vitro findings support the hypothesis that culture media supplemented with high concentrations of IGF-1 mixed in granulose cells and estrous cow serum can ameliorate the development of embryos maintained in vitro. IGF2, by contrast, is synthesized primarily in diverse tissues at the embryonic and fetal stages of mammalian growth. This work highlights the potential use of recombinant IGF-1 for the enhancement of embryonic development in cattle
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Arquivo Brasileiro de Medicina Veterinária e Zootecnia
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
