#TITLE_ALTERNATIVE#
The production rate of recombinant protein could be affected by presence of plasmid and <br /> <br /> composition of culture media. The presence of plasmid can be maintained with antibiotic <br /> <br /> selection markers on plasmid and addition of antibiotics into culture me...
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| Main Author: | |
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| Format: | Final Project |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/25629 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | The production rate of recombinant protein could be affected by presence of plasmid and <br />
<br />
composition of culture media. The presence of plasmid can be maintained with antibiotic <br />
<br />
selection markers on plasmid and addition of antibiotics into culture media. The regulatory <br />
<br />
agency restricted the uses of antibiotics in recombinant protein production due to safety reason. <br />
<br />
Culture media consists of several components such as peptone. Commonly used peptone is <br />
<br />
derived from animal protein, but currently soybeans derived peptone which is hydrolysed with <br />
<br />
papain is commercially available. This research was aimed to determine the effect of ampicillin <br />
<br />
and soy peptone on the production of superoxide dismutase recombinant Staphylococcus <br />
<br />
equorum (rMnSODSeq) in Escherichia coli TOP10. The rMnSODSeq encoding gene is carried by <br />
<br />
plasmid pCAD_sod_cer. The first step of this research is confirmation of recombinant E. coli that <br />
<br />
carry plasmid pCAD_sod_cer. The plasmid was isolated and confirmed by migration and restriction <br />
<br />
analysis. The confirmed E. coli was grown on Terrific Broth (TB) and Luria Bertani (LB) media using <br />
<br />
soy pepton or animal pepton with and without ampicillin. The effect of ampicillin and soy pepton <br />
<br />
on rMnSODSeq production were observed with SDS-PAGE and ImageJ software. The stability of <br />
<br />
plasmid segregation was determined by plasmid retention. Soy peptone increased rMnSODSeq <br />
<br />
production rate by 2.6 times compared to peptone from animals in TB and 1.4 times in LB with or <br />
<br />
without ampicillin. Plasmid pCAD_sod_cer has good segregation stability (> 90 %) even without <br />
<br />
ampicillin on media using soy peptone and pepton from animal. The use of ampicillin for the <br />
<br />
production of rMnSODSeq with pCAD_sod_cer can be removed and soy pepton can increase its <br />
<br />
production rate. <br />
<br />
<br />
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