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E.coli galk和gpt基因重组质粒(pIDB103)导入金鱼受精卵内的研究
引用本文:于建康,王亚平,黄河,杜淼,李光三,张玉廉,陆德裕.E.coli galk和gpt基因重组质粒(pIDB103)导入金鱼受精卵内的研究[J].中国生物化学与分子生物学报,1989,5(5):417-422.
作者姓名:于建康  王亚平  黄河  杜淼  李光三  张玉廉  陆德裕
作者单位:中国科学院发育生物学研究所 北京 (于建康,王亚平,黄河,杜淼,李光三,张玉廉),中国科学院发育生物学研究所 北京(陆德裕)
基金项目:美国洛氏基金会RF84031基金
摘    要:用显微注射法把含有E.coli galk和gpt基因的环状和线状重组DNApIDB103分别导入金鱼受精卵的细胞质内。这些注射过的卵子一般都能正常发育。从各不同发育时期的胚胎分离DNA与~(32)P标记的pIDB103探针杂交表明,导入的环状外源重组DNA在胚胎发育的早期,绝大部分以各种环状构型存在。从原肠胚晚期开始,它们逐渐形成串联状高分子量DNA。在尾芽期仍能检测到它们的序列。但尚未证明,它们是否与受体的染色体DNA发生整合。我们从囊胚期的胚胎中回收到了能转化大肠杆菌的环状重组DNA,它的酶切图谱和pIDB103极其相似。导入金鱼受精卵内的线状重组质粒pIDB103,除少量DNA与金鱼的染色体DNA可能发生整合外,其余绝大部分也形成高分子量DNA。

关 键 词:基因转移  分子杂交  
收稿时间:1989-10-20

Study of Eschcrichia coli Galk and Gpt Gene Introduced Into Fertilized Eggs of Goldfish
Yu,Jian-kang Wang,Ya-ping Huang,He Du,Miao Li,Guang-san Zhang,Yu-lian Lu De-yu.Study of Eschcrichia coli Galk and Gpt Gene Introduced Into Fertilized Eggs of Goldfish[J].Chinese Journal of Biochemistry and Molecular Biology,1989,5(5):417-422.
Authors:Yu  Jian-kang Wang  Ya-ping Huang  He Du  Miao Li  Guang-san Zhang  Yu-lian Lu De-yu
Institution:(Institute of Developmental Biology, Academia Sinica, Beijing
Abstract:Circular and linear plasmid pIDB 103 contained E. coli galk and gpt genes were microinjected into the cytoplasm of fertilized eggs of goldfish. The exogenous DNA sequences seems to be stable in early embryonic stages of the goldfish. At the begining of embryos injected with circular plasmid, the plasmid DNA sequences were detected as circular conformation by southern blot. From gestrula stage, some plasmd DNA sequences were detected as linear conformation with high molecular weight and comigrated with cellular DNA on agarose gel electrophoresis. The circular plasmid were rescued from the injected blastula by retrasformaing E. coli HB 101. Restriction endonuclease analysis of this DNA suggested that the majority of the injected circular DNA were not modified following replication in goldfish embryos. When fertilized eggs of goldfish injected with linear plasmid pIDB 103, the most exogenous DNA sequences were detected as linear conformation with high molecular weight. Some of which seems to be integrated into the chromosome of goldfish.
Keywords:Gene transfer Hybridization
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