噬菌体φHAU3的cos位点及其与寄主相互作用的功能位点attP和attB的定位

LOCALIZATION OF cos AND attP SITES ON PHAGE HAU3AND attB SITE ON THE HOST CHROMOSOMES

ＨＡＵ３是寄主范围很广的放线菌噬菌体。Ｓｏｕｔｈｅｒｎ杂交实验表明，ＨＡＵ３可以整合到吸水链霉菌应城变种１０－２２和变铅青链霉菌６６的突变体ＺＸ１的染色体中，形成溶原，其溶原菌自发释放ＨＡＵ３，不受热激和紫外线照射的诱导。通过比较ＨＡＵ３衍生噬粒ｐＩＪ８３００的ＤＮＡ酶切片段在加热前、后电泳带谱的区别，将ＨＡＵ３的ｃｏｓ位点在ｐＩＪ８３００的图谱上得到了定位。还利用Ｓｏｕｔｈｅｒｎ杂交的方法定位了ＨＡＵ３与宿主形成溶原时附着位点（ａｔｔＰ），并利用脉冲电泳技术定位了在变铅青链霉菌ＺＸ７和吸水链霉菌应城变种１０－２２中形成溶原的附着位点（ａｔｔＢ）。这些信息均有利于以ＨＡＵ３为基础的载体的发展和优化。

opwood Actinophage HAU3 has a wide host range. Southern hybridization indicates that HAU3 DNA can lysogenize Streptomyces hygroscopicus 10-22 as well as Streptomvceslividans ZX7 by integrating into their chromosomes.No increase of phage production was ob-served after UV irradiation or incubdtion of HAU3 lysogenic spores. Comparison of the fragmentpatterns on agarose gels of annealed and heated digests of HAU3 derivative,plasmidpIJ8300 DNA (isolated from phage particles),allowed us to localize the HAU3 cos site on therestriction map of plasmid pIJ8300. In addition,attP site of HAU3 and attB site on thechromosome of Streptomyces lividans ZX7 and Streptomyces hygroscopicus var.yingchengensis10-22 strains have also been located by the use of pulsed-field gel electrophoresis and Sou-them hybridziation. These informations will be helpful for the development of HAU3 touseful gene cloning vector(s).