Kul'ba A M, Gorelyshev A S, Evtushenkov A N, Fomichev Iu K
Mol Gen Mikrobiol Virusol. 1991 Dec(12):26-9.
The recombinant bacteriophages with the genomes containing the DNA fragments of bacteria Erwinia chrysanthemi, including the pectatelyase gene, were constructed on the base of Pseudomonas aeruginosa temperate bacteriophage SM. The gene transferred into Pseudomonas aeruginosa PAO1 cells by transfection is expressed in the new bacterial host. The restriction maps of the recombinant bacteriophages are constructed and the position of an insert is defined. Bacteriophage SM was found to be capable of reproducing in Pseudomonas aeruginosa PAO1 cells when its DNA was shortened to 88% or increased to 111% of the normal genome length. Except for bacteriophage SM, the recombinant bacteriophage SM-2 with an unique restriction endonuclease site for XbaI can also be used as a vector for cloning. Bacteriophage SM capacity in cloning of heterological DNA at HindIII sites is not less than 8 Md, capacity of bacteriophage SM-2 is not less than 5 and 8 Md at XbaI and HindIII sites respectively.
以铜绿假单胞菌温和噬菌体SM为基础构建了基因组包含菊欧文氏菌DNA片段(包括果胶酸裂解酶基因)的重组噬菌体。通过转染导入铜绿假单胞菌PAO1细胞的基因在新的细菌宿主中表达。构建了重组噬菌体的限制性图谱并确定了插入片段的位置。发现当噬菌体SM的DNA缩短至正常基因组长度的88%或增加至111%时,它能够在铜绿假单胞菌PAO1细胞中繁殖。除了噬菌体SM外,具有独特XbaI限制性内切酶位点的重组噬菌体SM-2也可作为克隆载体。噬菌体SM在HindIII位点克隆异源DNA的能力不小于8 Md,噬菌体SM-2在XbaI和HindIII位点的克隆能力分别不小于5 Md和8 Md。