摘要
以胡萝卜鱼雷形胚状体为材料，以λｇｔ２１０噬菌体为载体，构建了一个含有６．０×１０＾８个重组子的ｃＤＮＡ文库。用ＰＣＲ法扩增的长度为１．１ｋｂ的胚性细胞蛋白６３ＤＮＡ片段作探针，从ｃＤＮＡ文库中筛选出一个完整的ＥＣＰ６３ｃＤＮＡ克隆。
cDNA library with about 6.0×108 plaques per g phage from carrot (Daucus carota L.
cv. Harumaki) torpedo-shaped embryos was constructed by a cDNA cloning system (λgt10). A full-length cDNA for embryogenic cell protein 63 (ECP63), an embryogenic cell protein from carrot with a relative molecular weight of 63 000, was isolated from a cDNA library using PCR-amplified DNA as a probe. The nucleotide sequence of ECP63 cDNA was 1 989 bp in length. The cDNA encoded a polypeptide of 569 amino acids, and the calculated molecular weight of this polypeptide was 62 000. The Northern blot analysis using labeled full-length ECP63 cDNA as a probe showed that the gene for ECP63 was expressed in embryogenic cells,globular,heart-,and torpedo-shaped embryos,but not in seedlings and non-embryogenic cells.
基金
国家教委回国留学人员基金
关键词
胡萝卜
胚性细胞蛋白
胚状体
胚胎发生
基因
Daucus carota,, Embryogenic cell protein, Embiyoid, Embryogenesis, Gene
