Utilization of 14c Phenylalanine Derived from Arylphorin or Free Amino

UTILIZATION OF {14C} PHENYLALANINE DERIVED FROM ARYLPHORIN OR FREE AMINO ACID IN MANDUCA SEXTA PHARATE ADULTS GENERAL INTRODUCTION TO THE PAPER How did someone figure out that carbon dioxide in the bloodstream is exchanged for oxygen in the lungs of mammals? And how do we know that hormones are produced by, for example, the pituitary gland and, moreover, that these hormones signal the body to grow. These and many other questions have been answered by means of the scientific method. Scientists form hypotheses, experiment, collect data, and draw conclusions from this data that is then applied to answering questions about that certain topic. Some research seems to be obselete and/or practically useless, only to come into the picture years or decades later and shift the field of science. Although at times it is difficult to say what the repercussions of a particular study will be, it is not uncommon for that very study to play a key role in a major scientific breakthrough later. Therefore, it is important to keep in mind that although a particular study may not seem to be important at the time, later it could be the most signifi cant study in the field. This study performed by Min Wu and Dr. Marc E. Tischler, at first glance may appear to have a narrow focus, but upon furthur consideration, one realizes that its findings have applications to any number of related topics. In this experiment, Wu and Tischler tracked what paths a specific protein, arylophorin, and followed its production and distribution throughout the body of Manduca sexta pharate adults. Their first goal was to determine the most efficient method for producing radioactively labeled arylophorin, called 14C-Arylphorin. Their second goal, and purpose of the experime... ...test) difficulty of the experiment--finding the right conditions from which the highest (or most acceptable) specific activity could be achieved. After much experimentation it was determined that incubating one-half fat body in 1 ml of medium (0.15 mM of labeled phenylalanine and amino acids at 25% normal level in Grace's medium) for 15-24 hours, chasing labeled phenylalanine after one hour with unlabeled phenylalanine, yielded most desirable results. Temperature was shown to have little or no affect on the results, so room temperature was used in further experimentation and final implementation in achieving high specific activity (more convenient). REFERENCES Tischler, Marc E., Wu, Min. (1995) Utilization of 14C Phenylalanine Derived From Arylphorin or Free Amino Acid in Manduca sexta Pharate Adults. Archives of Insect Biochemistry and Physiology pgs.258-272.