Paper Information

Journal:   IRANIAN JOURNAL OF PUBLIC HEALTH   AUGUST 2014 , Volume 43 , Number SUPPLEMENT 2; Page(s) 45 To 45.
 
Paper: 

DESIGN AND CLONING OF RECOMBINANT PEPTIDE DRUG GENE, TERIPARATIDE IN E.COLI BL21 (DH5?)

 
 
Author(s):  SAGHARIDOOZ SEPIDE, BAKHTIARI NAHID
 
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Abstract: 

Background: The aim of this research is to design and to clone the rhPTH gene in E. coli BL21 (DE3).
Methods: This research discloses a novel frame designed for efficient preparation of N-terminal 1-34 amino acids of hPTH as functional parts of this hormone. The frame constructed as an NcoI-BamH1 fragment encoding a His-tag and a chimeric fusion protein consisting of a fusion partner comprising of 52 amino acids belonging to Escherichia coli
b- galactosidase (LacZ) gene, a cleavage site identified by Entropeptidase and rhPTH (1-34) gene fragment. Optimized frame was synthesized and ligated with pET28a vector under the control of T7 promoter, and then transformed in E. coli BL21 (DH5a) cells.
Results: Positive clones that released the mentioned frame by double digestion with NcoI-BamH1 enzymes were approved by sequencing.
Conclusion: We cloned designed gene in E. coli Bl21 (DH5
a). We expect to express our designed gene to be very elevated after itsinduction by IPTG in Escherichia coli BL21 (DE3).

 
Keyword(s): E. COLI, TERIPARATIDE, OSTEOPORESIS, RHPTH-CLONING
 
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