胡萝卜(Daucus carota L.)胚性细胞蛋白的分离研究

应用IEF/SDS-PAGE双向电泳技术,比较了胡萝卜胚性细胞、非胚性细胞和不同发育时期的胚状体中可溶性蛋白的双向电泳图谱,结果发现在胚性细胞中特异存在的胚性细胞蛋白在不同发育时期的胚状体中也存在,但在非胚性细胞中不存在。因此,推测体细胞胚胎发生所需的一些基因在胚性细胞中就早已表达了。我们还成功地分离和测定了ECP 45-2 N-末端和中央部分氨基酸序列。与已知氨基酸序列相比,ECP 45-2部分氨基酸序列与ECP 45-1 部分氨基酸序列具有较高比例的同源性。因此, ECP 45-1和45-2可能属于同一因基家族。

ISOLATION AND SEQUENCING OF EMBRYOGENIC CELL PROTEIN FROM CARROT

Total soluble proteins were extracted from non-embryogenic cells, embryogenic cells and different developmental stage of somatic embryos from carrot (Daucus ca-rota L.) and were analyzed by isoelectric focusing sodium dodecylsulfate-polyacry-lamide gel electrophoresis(IEF/SDS-PAGE) two dimensional electrophoresis. The results demonstrated that two spots of 63-65 KDa molecular weight with pI ranging from 6.2-6.0 and two spots of 45 KDa molecular weight with pI ranging from 5.2-5.0, which were detected in embryogenic cells and undetected in non-embryogenic cells, were also detected in the extracts of different somatic embryos (globular stage,heart-shaped stage and torpedo-shaped stage). We concluded that some genes necessary for somatic embryo formation are already expressed in embryogenic cells. We successfully isolated and determined the partial amino acid sequences of ECP 45-2, a carrot embryogenic cell protein. The partial amino acid sequences of ECP45-2 show higher homology to ECP 45-1, which suggests that the genes for ECP45-1 and 45-2 are to be consist of a same gene family.