Transcription of T7 DNA immobilised on latex beads and Langmuir-Blodgett film.
The recognition of DNA is the first and most important condition for biological applications, including transcription and translation regulators and DNA sensors. For this purpose, we have developed few systems where we were able to immobilize long double-stranded DNA (dsDNA) successfully to the surfaces of different solid substrates. To achieve this, we have chosen polystyrene beads and standard Langmuir-Blodgett monolayer of Zn-arachidate. In the first attempt, variant of T7 DNA containing one strong promoter A1 for Escherichia coli RNA polymerase was immobilised on uniform polystyrene microspheres (0.31 microm diameter) by covalent grafting. In the latter case, Zn(II) is bound to arachidic acid through charge neutralization. Since tetrahedral Zn(II) participates in DNA recognition through coordination, we have been able to layer DNA over the Zn-arachidate monolayer. The successful immobilization of DNAs on these different substrates was visualized under fluorescence microscope. These immobilized DNAs were used as a template to study in vitro transcription reaction and thus we introduce a new strategy for the study of transcription in heterogeneous phase.
Ghosh
Debalina
D
Molecular Biophysics Unit, Indian Institute of Science, Bangalore-560012, India.
Ramakanth
Madhugiri
M
Bhaumik
Anusarka
A
Faure
Nicolas
N
Rondelez
Francis
F
Chatterji
Dipankar
D
D000327:Adsorption; D017123:Bacteriophage T7 / Q000235:genetics*; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry; D004537:Eicosanoic Acids / Q000737:chemistry; D008567:Membranes, Artificial; D008863:Microspheres; D014158:Transcription, Genetic
D020099:Coated Materials, Biocompatible; D015342:DNA Probes; D004279:DNA, Viral; D004537:Eicosanoic Acids; D008567:Membranes, Artificial; D012321:DNA-Directed RNA Polymerases; C094477:arachidic acid