Construct recombination ibrary

We just demonstrated the feasibility of construction of library of recombination segments like the one in humans. In future work, we hope to finish this part and make real antibody libraries.

The protocol for molecular manipulation is clear but we may need a larger vector, such as the BACs to harbor all the segments.

Test of antibody function

Once the antibody library is accomplished, the major focus will be whether such antibodies can have normal function. We will settle such issues with antibody expression and subsequent assays. Immunohistochemistry or immunofluorescence offers the opportunity to compare the specificity of such antibodies while equilibrium dialysis can measure its affinity.

Specificity

Affinity

Other applications

The recombination method is useful in other aspects.

Nowadays, microbe based environmental monitoring is fast and real-time. However, certain phenomenon might be instant but reveal the hint for potential environmental hazards. In this case, our system might be quite useful, for recombination records the instant event permanently in the genome of the bacteria, just like a log. Thus, detection will discover the appearance of the hazards even if it's transient. For instance, the increase in phosphate concentration in some districts might be transient but will cause algae bloom in lakes downstream. With this recombination apparatus, the bacteria which have encountered such abnormal phosphate concentration can undergo recombination and record this event permanently. Culture and analysis will reveal such harmful events.

Besides, since the signal can be recorded firmly with recombination, it can be amplified during proliferation of the bacteria. This is crucial for detecting some weak yet important changes.