Studies On Electrophoresis Technique Of Testing Seed Genetic Purity Of Chinese Cabbage

The concentrating mechanism of AU-PAGE was studied by comparing the electrophoresis patterns and measuring the voltage gradient during electrophoresis. On the basis of AU-PAGE electrophoresis system, electrophoresis technique of seed protein for Chinese cabbage (Brassica campestris L.ssp. pekinesis) was established. The results were as follows:1. The concentrating mechanism of AU-PAGEThe pH(4.6-4.7) of extracting solution consisting of TEMED (4%) and HAC (12%) is closed to the isoelectric points of many seed protein, so the protein has few electric charges and slowly and stop on the surface of gel due to the gel barrier effect. In the meantime, TEMED move into gel slowly until the TEMED entered gel all and protein accumulated to a thin zone on the surface of gel.2. Role of TEMED on protein isolationTEMED entered gel reduces the voltage gradient in this region and remains a zone what has high voltage gradient, in which protein enter the gel and move faster but slower than TEMED. Different protein are separated according to their molecular mass and electric charge3. Electrophoresis system of Chinese cabbage seed proteinThe electrophoresis technique is as follows in detail.(1)Seed protein extracting: The seeds are smashed and put into the 0.5ml tube, then adding 40 l water in the tube for forty minutes, then centrifuging 3 minutes at 10000 rpm, take out 15 l supernatant which will be diluted 2 times with regulation solution that has 10%TEMED, 15%urea and 30%HAC.(2)Stank gel solution: Gel solution has 10%acrylamide, 0.25%Bis, 10% urea,2%HAC, 0.4%NaACand 0.02%FeSO4.(3) Electrophoresis: Electrophoresis is running in 2% acetic acid electrode solution at 40mA stability current per gel.4. Correlation analysis between electrophoresis test and plot testSeven samples purity of Chinese cabbage was tested with above system and in the field with plot test. Correlation analysis between the electrophoresis test (X) and plot test (Y) showed that there is significant relation between two methods and regression equationis Y=0.97X+4.35...