Difference between revisions of "Team:Stuttgart/Keratinases"
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<b>Figure 1:</b> Extracellular total protein concentration of three different keratinases (kerUS, kerA, kerP) each of them located in different vectors (pet28, pSB1C3 and pSB1K3). | <b>Figure 1:</b> Extracellular total protein concentration of three different keratinases (kerUS, kerA, kerP) each of them located in different vectors (pet28, pSB1C3 and pSB1K3). | ||
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Revision as of 23:29, 1 November 2017
Results
Keratinases
BCA Assay
Protein concentration of supernatant and cell disruption of E.coli carrying different keratinase plasmids (KerP, KerUS and KerA in different vectors) was measured with BCA Assay method (see chapter methods). The figure 1 below shows the BSA standard curve (blank corrected, arithmetic average of triplicates). Table 7 and 8 show the results of sample measurements (OD values: blank corrected, arithmetic average of triplicates). The concentrations are calculated with the calibration curve (BSA standard curve). Expectetly protein concentration is much more higher in cell disruption than in supernatants. It is also noticeable that protein concentraion of E.coli wild type is very high compared to engineered cultures. Unfortunately we do not have OD values, but we explain it by a higher amount of wild type cells after cultivation.
Table: supernatant (dilution 1:25):
| sample | OD562 | concentration [µg/mL] |
|---|---|---|
| KerP psD1K3 | 0,194 | 3066,667 |
| KerP psD1C3 | 0,195 | 3091,667 |
| KerUS pet28 | 0,191 | 2991,667 |
| KerUS psD1C3 | 0,207 | 3391,667 |
| KerUS BB | 0,226 | 3866,667 |
| KerA pet28 | 0,194 | 3083,333 |
| KerA psB1C3 | 0,213 | 3550,000 |
| KerA BB | 0,201 | 3241,667 |
| wild type E.coli | 0,248 | 4416,667 |
Table: cell disruption (dilution 1:25):
| sample | OD562 | concentration [µg/mL] |
|---|---|---|
| KerP psD1K3 | 0,514 | 11075,000 |
| KerP psD1C3 | 0,611 | 13491,667 |
| KerUS pet28 | 0,405 | 8358,333 |
| KerUS psD1C3 | 0,285 | 5350,000 |
| KerUS BB | 0,385 | 7841,667 |
| KerA pet28 | 0,482 | 10266,667 |
| KerA psB1C3 | 0,585 | 12858,333 |
| KerA BB | 0,410 | 8475,000 |
| wild type E.coli | 0,615 | 13600,000 |
HIER STEHT EIN TEXT
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FÜLL MICH MIT WORTEN
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