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促黄体素释放激素基因的化学合成与克隆
Chemical Synthesis and Cloning of Mammal and Avian Luteinizing Hormone Releasing Hormone Genes
【摘要】 用固相亚磷酰胺法合成了家畜和家禽的促黄体素释放激素基因的3个寡核昔酸(L1、L2和L3)片段。经退火,聚合成平端双链后,以BamHI和HindⅢ双酶切,插入载体pBluescript的多克隆位点中,通过菌落原位杂交、酶切位点分析及聚丙烯酞胺凝胶电泳鉴定,证实化学合成的促黄体素释放激素基因已在E.coliDH5a中克隆成功,序列分析结果表明人工合成的基因序列完全正确。
【Abstract】 The genes coding for mammal and avian luteinizing hormone releasing hormone(LHRH)havebeen chemically sythesized by solid-phase phosphoramidite methed. The 51 base-pair synthetic DNA duplexconsists of structural gene encoding LHRH,two stop codon TAA,TGA and Hind Ⅲ restriction site at 3’end and BamHI restriction site at 5’end of gene. The recombinant colonies were identified by in situ hy-bridization and restriction enzyme digestion,Their accuracy was comfirmed by DNA Sequence Analysis.
【关键词】 基因的化学合成;
促黄体素释放激素;
基因克隆;
【Key words】 chemical synthesis of the gene; luteinzing hormone releasing hormone; DNA cloning;
【Key words】 chemical synthesis of the gene; luteinzing hormone releasing hormone; DNA cloning;
【基金】 “863”计划生物技术领域青年基金
- 【文献出处】 农业生物技术学报 ,JOURNAL OF AGRICULTURAL BIOTECHNOLOGY , 编辑部邮箱 ,1994年01期
- 【分类号】S852.2
- 【被引频次】7
- 【下载频次】51