细胞分裂素的混合抗体型免疫亲和柱的制备与应用

Preparation and Application of the Mixed-antibody Immunoaffinity Column for Gytokinins

内源细胞分裂素可以分成三组：异戊烯基腺苷组（ｉＰＡｓ）、玉米素核苷组（ＺＲｓ）、二氢玉米素核苷组（ＤＨＺＲｓ）。从这三组分别选ｉＰＡ、ＺＲ、ＤＨＺＲ为半抗原合成免疫原，获得的三种免抗血清基本上只识别相应组的细胞分裂素。将经初纯化的三种抗血清偶联到ＣＮＢｒ活化的Ｓｅｐｈａｒｏｓｅ４Ｂ上制成免疫亲和柱。在其中的一根柱床高２．１ｃｍ、体积３．５５ｍｌ、直径１．５ｃｍ的亲和柱上，在流速１．５ｍｌ／ｍｉｎ、８０％冷甲醇为洗脱剂的条件下测得该柱容量为３．６—４．０μｇ、回收率达９３．９％～９８．３％。用该柱对丝瓜茎木质部伤流中细胞分裂素进行了纯化．表明能够用此柱对植物粗提液中的细胞分裂素进行快速分离纯化。

Endogenous cytokinins could be classified into three groups, that is, isopentenyl adenosine-group (iPAs ), zeatinriboside-group (ZRs ): and dihydrozeatinriboside -group (DHZRs ). Three haptens (iPA, ZR.and DHZR) have been. selected and the three respective im munogens have been synthesized.Three kins of antisera have been raised in raabbits and each kind onlyirecognizes therelative group (iPA-gronp,ZR-group,or DHAR-group cytokinins).After par tialpruication,these three kinds of antisera have been coupled to the CNBractivated Sepharose 4B.Then through the mixed-antibody immunoaffinity column the cytokinins contained in plant extracts could be fast and almost throughly isolated and pruified .This column is characteristic of the bed height of 2.1 cm ,the bed volume of 3.55 ml,the bed diameter of 1.5cm And the dey measuremen parameters have been determined as the column capacity of 3.6-4.0ng,the capture effieiency of 93 9%-98.3%.and the flow rate of 1.5ml/min.80% cold methanol has been chosen ad the eluent and phosphate buffer salt (10mmol/L,ph 7.4)as the recovery solution,With this column,cythkinins in xylem saps from stems of the towel gourds have been rapidly purified.