DNA interpolyelectrolyte complexes as a tool for efficient cell transformation.
A tool was developed for enhancement of plasmid penetration into an intact cell, based on increasing DNA hydrophobicity via inclusion into a soluble interpolyelectrolyte complex (IPC) with polycations. The characteristics of formation of DNA IPC with synthetic polycations [poly(N-ethyl-4-vinylpyridinium)bromide (PVP) and PVP modified with 3% of N-cetyl-4-vinylpyridinium units (PVP-C)] were studied using ultracentrifugation and polyacrylamide gel electrophoresis methods. The conditions were established under which the mixing of DNA and polycation aqueous solutions results in the self-assembly of soluble IPC species. Incorporation of DNA into IPC results in the enhancement of DNA binding with isolated Bacillus subtilis membranes. A considerable increase in the efficiency of transformation of B. subtilis cells with pBC16 plasmid resulted from incorporation of the plasmid into the IPC with PVP and CVP.
Kabanov
A V
AV
Research Center of Molecular Diagnostics, USSR Ministry of Health, Moscow.
Astafyeva
I V
IV
Chikindas
M L
ML
Rosenblat
G F
GF
Kiselev
V I
VI
Severin
E S
ES
Kabanov
V A
VA
D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism; D010582:Bacteriophage lambda / Q000235:genetics; D002462:Cell Membrane / Q000378:metabolism; D004269:DNA, Bacterial / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D004279:DNA, Viral / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000187:drug effects*; D011145:Polyvinyls / Q000494:pharmacology*; D014169:Transformation, Bacterial; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D004279:DNA, Viral; D011145:Polyvinyls; C027968:poly-4-vinyl-N-ethylpyridine