Amplification, cloning and expression of the Reg3 a gene from mouse pancreas. | [Pakistan Journal of Biotechnology • 2015]

Author(s):

1. Tehmina Siddique: Diabetes and Cardio-Metabolic Disorders Laboratory, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box-577, Faisalabad, Pakistan

2. Fazli Rabbi Awan: Diabetes and Cardio-Metabolic Disorders Laboratory, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box-577, Faisalabad, Pakistan

3. Syeda Sadia Najam: Diabetes and Cardio-Metabolic Disorders Laboratory, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box-577, Faisalabad, Pakistan

4. Abdul Rehman Khan: Diabetes and Cardio-Metabolic Disorders Laboratory, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box-577, Faisalabad, Pakistan

5. Javed Anver Qureshi: Diabetes and Cardio-Metabolic Disorders Laboratory, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box-577, Faisalabad, Pakistan

6. Mohsin Khurshid: Diabetes and Cardio-Metabolic Disorders Laboratory, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box-577, Faisalabad, Pakistan

7. Mehboob Islam: Diabetes and Cardio-Metabolic Disorders Laboratory, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box-577, Faisalabad, Pakistan

8. Maryam Zain: Diabetes and Cardio-Metabolic Disorders Laboratory, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box-577, Faisalabad, Pakistan

Abstract:

Introduction: Reg proteins are a group of regenerating proteins which are implicated in the pancreas developmental biology. The aim of the study was to explore suitable conditions for cloning and expression of this recombinant protein that can ultimately be explored in further studies for testing its potential as a promising therapeutic for diabetes by inducing regeneration/neogenesis of pancreatic beta cells. Methodology: Total RNA was isolated from BALB/C mouse pancreas, cDNA was synthesized and Reg3 6 gene was amplified with gene specific primers. A recombinant plasmid (pET28a vector) was constructed with Reg3 6 gene. Recombinant protein was expressed in BL21 (DE3) strain in LB media. Expressed protein was isolated and separated on 12% SDS-PAGE. Results: Clones were confirmed with restriction and colony PCR and SDS-PAGE confirmed the 16 kDa band in IPTG induced samples. Further work for the purification of this protein will be pursued in future.

Page(s): 55-61

DOI: DOI not available

Published: Journal: Pakistan Journal of Biotechnology, Volume: 12, Issue: 1, Year: 2015

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