中国生物工程杂志

Protein extracting method、protein dissolution、protein loading amount and gel strip transfer were tested for the protein extracted from seedling rhizome of winter wheat cultivar Dongnong Winter Wheat No.1. The results were showed that the loss of low abundance protein in TCA/acetone protein extracting process (T process) was less than that in urea/thiourea protein extracting process (N process) and the spots in protein map in T process were more than that in N process. The purer protein dissolved two times in the hydrated liquid could maintain higher voltage of 8000 volt by Isoelectric Focusing and Electrophoresis. The protein loading of 10mg by two times dissolution in hydrated liquid could obtain clear map with more protein spots and better separation effect. After adding 400ul 0.3% common agarose solution to glue surface and washing support film of gel strip with 200ul electric buffer, the gel strips could be transferred easily to second dimension glue surface and no air bubbles appeared between gel strips and glue surface.