ASHLY+O.


 * __ Protein Expression Results: __**




 * Figure 1** experimental plate (Amp, BL21(De3), pGEM-gbr22) of N.R and C.T approximately 15 hours after being stored in 37 degree Celcius incubator (used for DNA sampling because growth was evident)




 * Figure 2** control plate (Amp, BL21(De3), No DNA), experimental plate (Amp, BL21(De3), pGEM-gbr22) and fun plate (no antibiotic, BL21 (DE3)) approximately 15 hours after being stored in 37 degree Celcius incubator


 * Figure 3** 50 ml conical tube with LB + amp, BL21(DE3), pGEM-gbr22 with purple wet pellet created after centrifuged for 10 minutes on 5,000 rpm in 4 degrees Celsius




 * Figure 4** flask of LB+Amp, BL21 (DE3) and pGEM-gbr22 after culture was incubated in 37 degrees Celcuis for 16-24 hours


 * __ Protein Purification Results: __**



=

 * Figure 1 ** 15 mL conical tubes with purified gbr-22 after being eluded from Ni-NTA resin by using a buffer with a high concentration of imidazole, Elution #1 after one wash and Elution #2 after a second wash =====




 * Figure 1a and 1b** wavelength (nm) versus absorbance (10 mm) for Elution 1 of pgbr-22, 1a absorbance = 0.54 mg/mL (1 Abs = 1 mg/mL) at 280 nm, 1b absorbance = 0.58 mg/mL (1 Abs = 1 mg/mL) at 280 nm, avg absorbance = .56 mg/mL


 * Figure 2a and 2b** wavelength (nm) versus absorbance (10 mm) for Elution 2 of pgbr-22, 2a absorbance = 0.21 mg/mL (1 Abs = 1 mg/mL) at 280 nm, 2b absorbance = 0.20 mg/mL (1 Abs = 1 mg/mL) at 280 nm, avg absorbance = .205 mg/mL
 * Figure 3a and 3b** wavelength (nm) versus absorbance (10 mm) for Elution 1 of pgbr-22, 3a absorbance = 0.110 mg/mL (1 Abs = 1 mg/mL) at 574 nm and .069 mg/mL (1 Abs = 1 mg/mL) at 280 nm, 3b absorbance =0.108 mg/mL (1 Abs = 1 mg/mL) at 574 nm 0.062 mg/mL (1 Abs = 1 mg/mL) at 280 nm, avg absorbance at 574 = .109 mg/mL
 * Figure 4a and 4b** wavelength (nm) versus absorbance (10 mm) for Elution 2 of pgbr-22, 4a absorbance = 0.018 mg/mL (1 Abs = 1 mg/mL) at 574 nm and .021 mg/mL (1 Abs = 1 mg/mL) at 280 nm, 4b absorbance = 0.018 mg/mL (1 Abs = 1 mg/mL) at 574 nm and 0.021 mg/mL (1 Abs = 1 mg/mL) at 280 nm, avg absorbance at 574 = .018 mg/mL

__**Protein Characterization Results:**__
 * Figure 1** Protein characterization gel electrophoreses of pgbr-22 after gel was stained with bromophenol blue overnight and dried for 1.5 hours, lane 1 is cell lysate, lane 2 is the protein ladder, lane 3 is void, lane 4 is the soluble fraction, lane 5 is the flow through, lane 6 is the wash, lane 7 is elution 1 and lane 8 is elution 2, estimated protein molecular weight using ladder is ~25kDa

**Figure 2** PageRuler Prestained Protein ladder used as the molecular weight standard for purified gbr-22 protein