J Integr Plant Biol. ›› 1997, Vol. 39 ›› Issue (5): -.

• Research Articles •    

Purification of the Specific Plasma Membrane Proteins from the Surface of Sperm Cells of Zea mays

Yang Zhong-han, Zhang Yi-hong and Cao Zong-xun (Tsao T H)   

Abstract: Pollen samples of 6 varieties of Zea mays L. were used to isolate the viable sperm cells. After being probed with N-hydroxysuccinimido-biotin (NHS-biotin), the sperm cell plasma membrane proteins were compared with each other using the method of Western blotting. Results showed that there was no significant difference among varieties. The molecular weights of probed plasma membrane proteins were concentrated on 91,60,43,30 and 17 kD. Immunochemical method was adopted for further purification of sperm plasma membrane protein preparation which was some- what contaminated with cell organelles. After the cell organelles were isolated from etiolated seedlings of Zea mays by sucrose density gradient super centrifugation, the crude membrane proteins of organelles, endoplasm reticulum, mitochondria, Golgi body and plasmolemma were respectively used as antigen to immunize Guinea pig. The antibody was obtained from respective antiserum, then further used to produce immuno-affinity absorbent. After the solution of membrane proteins of sperm cells passed through the column, some proteins probed whth NHS-biotin were identified. Two major proteins probed with NHS-biotin were considered to be sperm cell specific. The size of these proteins in SDS-PAGE was about 65 kD, 22 kD, respectively.

Key words: Zea mays, Plasma membrane protein of sperm cells, Immuno-affinity chromatography, N-hydroxysuccinimido-biotin, Western blotting

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