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水稻叶绿体DNA的提取和纯化

Isolation and Purification of Rice Chloroplast DNA

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【作者】 沈志伟; 孙崇荣; 朱俭;

【Author】 Shen, Zhi-wei Sun, Chong-rong Zhu, Jian (Department of Biochemistry, Fudan University, Shanghai)

【机构】 复旦大学生物化学系; 复旦大学生物化学系 上海; 上海; 上海;

【摘要】 以籼稻品种珍讪97B为材料,采用溶液捣碎和不连续蔗糖梯度离心的方法提取了籼稻的叶绿体DNA,DNA经限制性内切酶酶解和琼脂糖胶电泳可以得到清晰的条带,来自蚕豆的核酮糖—1,5—二磷酸羧化氧合酶大亚基基因探针和23SrRNA基因探针可以与酶切条带杂交,由此确定了含这二种基因的BamHI酶切片段。

【Abstract】 The chloroplast was isolated and purified from the rice (Oryza Staiva, subsp. Indica, cv. Zhenshan 97B) leaves with liquid homogenizing and stepwise 35-55% sucrose gradient centrifugation. The pure chloroplast DNA (ctDNA) was isolated from it. The ctDNA was digested by several restriction enzymes and fragments were fractionated by agarose gel electrophoresis. The genes of rbcL and 23srRNA were located on the fragments cleaved by BamHI by broad bean chloroplast DNAs probes containing these genes.

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