Solenberg P J, Baltz R H
Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285.
Gene. 1994 Sep 15;147(1):47-54. doi: 10.1016/0378-1119(94)90037-x.
Transposons derived from the Streptomyces lividans insertion sequence IS493 are useful for the genetic analysis and manipulation of a number of Streptomyces spp. Tn5099-10, an IS493 derivative that contains a spontaneous deletion terminating in the left inverted repeat (IR-L), transposed at a 1000-fold higher frequency in Streptomyces griseofuscus, and at a tenfold higher frequency in Streptomyces fradiae, than the IS493 derivatives, Tn5096 and Tn5099. The IR-L from Tn5099-10 was used to construct a cassette which hypertransposes from plasmids containing the transposon genes, ORFA and ORFB, outside of the inverted repeats. The target sequences of two Tn5099-10 insertions conformed to the consensus target sequence of the other IS493 derivatives, gNCaNTgNNy (where lower-case letters indicate that other nt have been observed at this position and N is any nt). Transposition mutant libraries of S. griseofuscus and S. fradiae can be easily prepared in broth culture by using the hypertransposing elements and a temperature-sensitive delivery plasmid.
源自淡紫灰链霉菌插入序列IS493的转座子，可用于多种链霉菌属细菌的遗传分析和操作。Tn5099 - 10是一种IS493衍生物，含有一个终止于左侧反向重复序列（IR - L）的自发缺失，其在灰褐链霉菌中的转座频率比IS493衍生物Tn5096和Tn5099高1000倍，在弗氏链霉菌中的转座频率高10倍。来自Tn5099 - 10的IR - L被用于构建一个盒式结构，该结构可从含有位于反向重复序列之外的转座子基因ORFA和ORFB的质粒上进行高效转座。两个Tn5099 - 10插入的靶序列符合其他IS493衍生物的共有靶序列gNCaNTgNNy（其中小写字母表示在该位置观察到了其他核苷酸，N为任意核苷酸）。通过使用高效转座元件和温度敏感型传递质粒，可在肉汤培养中轻松制备灰褐链霉菌和弗氏链霉菌的转座突变体文库。
