Ion Exchange Fractionation of Rabbits Seminal Fluid: Recognizing a DNA Retardation Activity from the Main DNase Activity
The role of seminal proteins charge on the nature of seminal fluid inhibitory effect that exerted against exogenous DNA. Has been identified and an approach closely with more details to the nature of inhibitory activities available in rabbit seminal fluid proteins that prevent the entry of exogenous DNA into the head of sperm. After collection of rabbit’s ejaculate and removing sperm cells, seminal fluid was incubated with fixed concentration of exogenous DNA. The seminal fluid – exogenous DNA mixture was analyzed by electrophoresis. Ion exchange chromatography was used to separate seminal proteins on the basis of their charge. Positively charged proteins were eluted, lyophilized, and their profile was characterized by SDS-PAGE and native-PAGE. After incubation of this eluted group with the same source of DNA, the same electrophoretic conditions were applied on this group. According to our knowledge, this is the first paper in which ion exchange chromatography was used to separate two DNA counterfeiting activities of the seminal fluid using non-radioactive method in rabbits and even in other mammals. Thus, more than one inhibitory activity were identified and separated. DNA retardation activity (or DNA binding activity) which repressed DNA electrophoretic migration was the only activity that found to be available on the positively charged fractions while the DNase activity was found exclusively on the negatively charged group. Key words : Seminal fluid , transgenesis techniques , DNA electrophoretic migration , DNase