| ABSTRACT Some improvement in LEF/SDS-PAGE was made in this paper and the results were as follows: The cathode drift of IEF was restrained sharply when extreme nonequilibrum electrophoresis buffer was introduced to the isoelectrofocusing and the distribution of pH gradient in LEE gel was improved at the same time. Another way to extend the pH gradient distribution was the pH regulation of the sample bufi~r and sample overlay buffer of LEE properly. In this modified LEE protocol, a hermetic isoelectrofocusing system was set up to inhibit the CO2 absorption and, in the long run, the cathode drift was also restrainecL A high-volt-endured isoelectrofocusing system was established to enhance the differentiation ability of lEE. Furthermore, a new compensatory protocol of silver stain was also established in this study. In conclusion, all the patterns of LEF/SDS-PAGE showed that the differentiation power of the modified LEE protocol was improved greatly to meet the needs from ordinary biochemistry labs. The modified method was applied to analyze the proteins of thermosensitive genic male sterile rice(TGMSR) during the sensitive period of fertility regulation. It was found that only a few young panicle protein changes were detected and most of proteins kept the same. There were two kinds of changed protein spots in the TGMSR during the sensitive stage. One of them was related to the development of young panicle. It was common that some protein spots disappeared in the development of young panicle. E.g. 6 protein spots of Anxiang S disappeared at the 5th stage of young panicle development. Their distribution of molecular weight and p1 varied widely. And at the same development stage, 7 protein spots (MW 50-65k]), p1 5.0-6.1) which never appeared in the 4th ~tage of young panicle development were detected. At the later fertility sensitive period, 8 new protein spots compared with the 5th development stage were also found. Among them, the spot (MW lOkD, p1 6.48) was one of the most visible changed protein spots. But in Pei抋i 645, no protein spot related to the development stage of young panicle was detected. The other kind of protein change was related to the fertility regulation of TGMSR. Four kinds of protein change were found in this study. (1). Some protein spots disappeared only at the fertile temperature (22t) in the thermo-sensitive genic male sterile rice. In Anxiang 5, 32 protein spots, which distributed in the area of MW 10-31k]), p14.0-6.5, MW 32-45, p14.6-7.5 and near the MW 45kD, p1 4.3, disappeared at the fertile temperature. In Pei抋i 645, 13 protein spots disappeared during the early fertility sensitive period and 12 protein spots of lower molecular weight did under fertile condition. in I#眫4~it~ IEF/SDS-PAGE ~ (2). Some protein spots disappeared at the sterile temperature (>25C). 4 protein (MWI5-55kD) spots in Anxiang S disappeared under sterile condition and 1 protein spot did in the Pei抋i 645. it seemed that this kind of protein changes was less frequent than the protein changes at the fertile temperature. (3). The special proteins expressed only at the fertile temperature. In this study, only 1 protein spot was detected in the Anxiang S and none was found in the Pei抋i 64S. (4). The proteins only expressing at the sterile temperature were also found in Anxiang S (1 spot at the early fertility sensitive stage and 6 spots at the later st...
|
PDF Full Text Request