A Preliminary Study On The Proteome Of Rana Spinosa Muscle Between Tadpole And Frog

Rana spinosa is a kind of large economic frog which mainly lives in mountain districts of southern China. As it is of particular diet and economic values its proteomic research has enormous meanings.[Objective] Employing proteomic approaches, the protein expression levels have been compared by resovling all soluble and perimembrane proteins on the two-dimensional electrophoresis and the found different expressed proteins have been identified by mass spectrometry technology. As the proteins expressed differently in the muscle of Rana spinosa and its tadpole, they may have kind of relationship with metamorphosis, present study is considered to be of learned value on the animal auxanology.[Method] Present proteomic study is based on two-dimensional electrophoresis tandem mass spectrometry and other technologies. The proteins were extracted by mixed samples and one step method . The concentration of extracted proteins were determined by Bradford method. The same amount of proteins were applied on to the immobilized pH gradient strips (IPG strip pH3-10NL) and the total Vh( the product of voltage and hours) is 80kVh-85kVh . After two steps equilibrium, the first dimension IPG was applied on to the second dimension SDS-PAGE (13%, 1.5mm . Resulting gels were stained by commassie brilliant blue or silver which is in accordance with the applied mount of proteins . The images of two-dimensional electrophoresis were obtained by ImageScanner optical density scaner and further analyzed by PDQuest software. Some of the different expressed proteins were identified by tandem mass spectrometry (ABI 4700 TOF-TOF) using mascot software searching against NCBInr database. [ Result] The protein concentration of tadpole and frog are 11.4084μg/ul and 10.9956μg/ul respectively . There are 42 ± 5 different protein points in total. Ten of them were identified sucessfully:P1 is troponin T3, P2, P3, P6, P7are fast troponin T, P4 is Aldh-A protein, P5 is creatine kinase (EC2.7.3.2)-African clawed frog(fragment),P8 is Enigma-prov protein,P9, P10 are Ckm-prov protein.[Conclusion] (1) the protein extraction procedures established in this experiment can improve the extraction sufficiency of samples and the consequent resolving results are very...