RNase one gene isolation, expression, and affinity purification models research experimental progression and...

Area of Science:

• Biochemistry Laboratory Education
• Molecular Biology Techniques
• Enzyme Kinetics

Background:

• Traditional biochemistry labs often lack inquiry-based components.
• There is a need for structured yet flexible laboratory courses.
• Integrating gene expression and enzyme characterization provides a cohesive learning experience.

Purpose of the Study:

• To develop a biochemistry laboratory course centered on guided inquiry.
• To provide students with practical experience in molecular biology and enzymology.
• To foster scientific communication skills through presentations and poster sessions.

Main Methods:

• Isolation of the RNase One gene from prokaryotic genomic DNA.
• Expression of the RNase One gene as a tagged protein.
• Affinity purification of the expressed protein.
• Enzyme activity and substrate specificity assays.
• Student-led explanations of experimental procedures.

Main Results:

• Successful isolation, expression, and purification of RNase One.
• Characterization of enzyme activity and substrate specificity.
• Students developed detailed experimental explanations.
• A culminating poster session showcased student work.

Conclusions:

• The course successfully integrates instructor-guided and student-designed experiments.
• This approach provides a manageable entry into inquiry-based learning in biochemistry.
• The curriculum offers a cohesive narrative for diverse experimental activities.
• Students gain comprehensive skills in molecular biology and biochemical analysis.