[Cloning of large imperfect palindromes in circular and linear vectors].
Data on comparative analysis of cloning of large imperfect artificial palindromes and one natural palindrome in the circular pUC19 and linear pN15L vectors, constructed on the basis of temperate N15 bacteriophage minireplicon are presented. The artificial palindromes consisted of the two head-to-head oriented 5:5-kb lambda bacteriophage DNA fragments interrupted by a short sequence of varied size. Natural palindrome was represented by the 12.5-kb BamHI fragment of Tetrahymena pyriformis ribosomal genes cluster. Integration of some artificial palindromes and a natural palindrome into a circular vector resulted in a considerable decrease of its copy number. This was assumed to result from cruciform formation mediated by supercoiling of a circular vector DNA. Thus, a linear is vector preferable in cloning of inverted repeated DNA sequences.
Ravin
N V
NV
Bioengineering Center, Russian Academy of Sciences, Moscow, Russia.
Ravin
V K
VK
D000818:Animals; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D005822:Genetic Vectors; D007613:Kanamycin Resistance / Q000235:genetics*; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000235:genetics; D012093:Replicon; D015183:Restriction Mapping; D013604:T-Phages / Q000235:genetics*; D013769:Tetrahymena pyriformis / Q000235:genetics
Klonirovanie krypnykh neideal'nykh palindromov v kol'tsevom i lineĭnom vektorakh.