枯草芽孢杆菌噬菌体载体的构建

以φ0105DI：It为原始株构建的重组噬菌体φ105S35和φ10 5S36具有自主侵染能力和溶源化特征。其基因组内插入的lkb片段上的cat，基因赋予二者所在宿主以氯霉素抗性，在两株噬菌体中插入位点相同，即原φ105DI ：It的smal酶切片段D、E之间，但插入片段在二者中的定向相反。与cat基因同时引入的单一BamHI和Xbal位点提供了外源DNA的插入位置。重组噬菌体DNA可高效转染枯草芽孢杆菌原生质体。因此φ105S35和币φ105S36可作为枯草芽孢杆随系统的载体而被利用。

Construction of phage vector for Bacillus subtilis]

Recombinant phages phi 105S35 and phi 105S36, derived from phage phi 105DI:1t, remain the ability to infect and be lysogenized in the host cell. In the their genome there is a 1 kb inset fragment which occurs between the DNA fragments D and E resulted from SmaI degested DNA of phi 105DI:1t. But the orientation of this insert fragment is opposite in two recombinant phages, respectively. The cat gene on this fragment makes the host cell resistant to chloramphenaical. The unique BamHI and XbaI sites which were introduced into the genome of the recombinant phages provide inserting sites of foreign DNA fragment. DNA from these two recombinant phages can transfect protoplast of Bacillus subtilis at high frequency. Therefore, phi 105S35 and phi 105S36 are suitable as phage vectors in the molecular cloning of Bacillus subtilis.