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反向PCR方法克隆腾冲嗜酸两面菌的分子伴侣基因

Qing MA,Yu-ying ZHANG

MICROBIOLOGY（2005）

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Abstract
根据已知的Sulfolobus属的分子伴侣基因,设计简并引物,用PCR的方法从腾冲嗜酸两面菌(Acidianus tengchongensis)基因组DNA中分别克隆到了分子伴侣α亚基和β亚基的约500bp的基因片段.以它们为探针进行Southern杂交,确定了合适的限制性内切酶.以确定的限制性内切酶消化的基因组DNA环化物为模板,进行反向PCR反应,引物的延伸方向由已知序列出发沿环化分子向未知区域进行,扩增产物经测序表明为α亚基和β亚基基因.根据所得序列分别设计两对引物进行PCR,测序结果表明得到了α亚基和β亚基的完整基因.
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