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

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.Up to now the proteomic research of it has not been reported.[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 kidney of Rana spinosa and 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 strit 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 Umax scaner and furthedly 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 frog and tadpole is 11.96992μg/μl and 7.39515μg/μl respectively . There are 217 different proteinpoints in total. Two of them were identified sucessfully: Fpl is Gastrula zinc finger protein XLCGF42 and Tpl is nucleoside diphosphate kinase .[Conclusion] (1) the protein extraction procedures established in this experiment can improve the extraction sufficiency of samples and the consequent resolving results are very good. (2) IPG can increase the resolving power and repetition of two-dimensional electrophoresis . (3) The protein expression profiles of kidney between Rana spinosa and tadpole is very different . (4) Gastrula zinc finger protein XLCGF42 and nucleoside diphosphate kinase may have relationships with gene transcription and their sequences and functions ask for further research.