Alice Kirega 
University of New Hampshire
Major: Biology
Mentor: Dr. Stephen Torosian, UNH Department of Microbiology
Cloning Interleukin-2 Using Bacteriophage Lambda and Escherichia coli
Cloning of interleukin-2 has been a successful procedure in treating immune deficiency diseases such as cancer. Much is known about interleukin inducing activities, but its mechanism of function is poorly understood. Interleukin-2 are eukaryote cytokines protein regulators secreted by white blood cells and mainly function as growth factor to stimulate proliferation of immune cells, such as B and T-cells. IL play an important role in directing both innate and adaptive immune responses. The significance of cloning IL-2 in this project is to measure whether the cloning will be successful through lambda vector.
Prior to the cloning of IL-2, a single stranded IL-2 RNA must be converted to double stranded DNA through reverse transcriptase PCR. After this conversion, the IL-2 DNA is inserted in a vector (lambda), to make a cloned DNA. This cloned DNA is then exposed to a host cell, E. coli, so that the cloned DNA can diffuse into the host cell membrane through electroporation. When the cloned DNA diffuses in a host cell, it integrates with the host genome therefore it's replicated along with host DNA. The phage clones are released as E. coli lyses.
Escherichia coli is a bacteria infected by a bacteriophage in nature, and commonly used as a host in cloning. Cloning using bacteriophage lambda as a vector is a well practiced procedure in microbiology because the structure of lambda's capsule makes integrating and packaging foreign DNA more proficient. Continuous cloning of interleukin genes can help to increase our knowledge about its specific interaction with immune functions. This will help us to develop a good approach to treat viral immune infections.
