摘要
放线菌噬菌体φHAU3是以吸水链霉菌应城变种10-22为指示菌,筛选获得的一个广谱性噬菌体,其基因组大小约为53kb,具有粘性末端。已成功地将44.7kb的φHAU3 DNA克隆到pIJ255上,构建成噬粒(Phasmid)pIJ8300;46.8kb的φHAU3 DNA克隆到pIJ285上,构建成pIJ8301。采用部分酶解和Southern杂交的方法,制作了pIJ8300和pIJ8301被Asp718和Bgl Ⅱ酶切的限制酶图谱,又综合单,双和多酶切分析的结果,定位了单一的Pvu Ⅱ和NcoI切点。综合噬粒pIJ8300和pIJ8301的特性,已将φHAU3中噬菌体功能的必需区定位在41kb的范围内。这些结果为以φHAU3为母体发展新的放线菌噬菌体奠定了良好的基础。
φHAU3 is a wide host-range actinophage isolated from soil on Stre-ptomyces hygroscopicus 10-22 as indicator. The genome of φHAU3 is about 53 kb long with cohesive ends. Two phasmids, pIJ8300 and pIJ8301, were constructed when part of the φHAU3 DNA were cloned into two cosmid vectors, pIJ255 and pIJ285, respectively. pIJ8300 carries 44.7 kb φHAU3 DNA in pIJ255, whereas pIJ8301 carries 46.8 kb DNA in pIJ285. The restriction maps of pIJ8300 and pIJ8301 with sites for Asp718, Bgl Ⅱand unique sites for PvuⅡ and Ncol had been constructed using the method of partial digestion and Southern hybridization. The combination of the properties of pIJ8300 and pIJ8301 allowed localization on φHAU3 the 41 kb essential region for plaque formation in Streptomyces, which should be very helpful for the development of new phage cloning vectors based on φHAU3.
出处
《病毒学报》
CAS
CSCD
北大核心
1993年第4期367-373,共7页
Chinese Journal of Virology
基金
国家自然科学基金
关键词
放线菌
噬菌体
噬菌斑
基因克隆
Actinophage, Gene cloning vector, Restriction mapping, Southern hybridization
