Stuchlík S, Janitorová V, Turna J
Department of Molecular Biology, Comenius University, Faculty of Natural Sciences, Bratislava, Slovak Republic.
Acta Virol. 1993 Oct;37(5):369-76.
We describe an in vivo cloning method using mini-Mu phage for genes, which cannot be cloned on multicopy vectors, mainly for their toxicity. We have successfully cloned succinate dehydrogenase (sdh) gene E. coli which was inactivated with defined insertion of fragment Kmr by this method. The most of obtained Kmr clones of mini-Mu transductants have contained the sequence of whole sdh gene. The intact gene of sdh can be reconstructed by site-directed mutagenesis.
我们描述了一种利用微型 Mu 噬菌体进行体内基因克隆的方法，该方法主要用于克隆那些因毒性而无法在多拷贝载体上克隆的基因。我们已成功利用此方法克隆了大肠杆菌的琥珀酸脱氢酶（sdh）基因，该基因通过特定片段 Kmr 的插入而失活。大多数通过微型 Mu 转导获得的 Kmr 克隆都包含完整的 sdh 基因序列。完整的 sdh 基因可通过定点诱变进行重建。
