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Construction of New Recombinant Plasmid Carrying Bone Morphogenetic Protein Gene

Rahimi Zarchi, Mahmood | 2012

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 43544 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Yaghmaei, Soheyla; Mashayekhan, Shohreh; Khoshzaban, Ahad
  7. Abstract:
  8. The purpose of this reaserch is to create a context in production of Bone Morphogenetic Protein-2 with more effective properties in Bone healing in Iran. Because of demand Of this protein in medical field it could cause an evalution in bone healing. The first section of this reaserch was the Construction of new recombinant plasmid carrying bone morphogenetic protein gene that was title of this thesis. First total RNA was extracted from ostosarcoma cancer cell line MG-63 followed by cDNA synthesis.For amplifying of the mature region of protein a pair of primer with tow restriction site was designed. After Success in PCR and obtaining PCR product with size of 343bp it transferd to vector Pet32a+ . The recombinant vector was transferred to the host which was E.coli strain BL21 DE3. Then the bacteria were cultivated in a liquid medium containing antibiotic and specialized marker for screening. After extracting the bacteria containing the recombinant vector , their non-genomic plasmid were extracted.The PCR test of this plasmid had a 343 bp PCR product that shows they are recombinant plasmid.This stage is a prerequisite for expressing and down stream processes for purification and activation of protein.
     
  9. Keywords:
  10. Gene Cloning ; Recombinant Plasmia ; Bone Morphogenetic Proteins

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