A novel approach to design of cis-acting DNA structural element for regulation of gene expression in vivo

Sarkar, Parthas S and Bagga, Rajesh and Balagurumoorthy , P and Bramachari, Samir K (1991) A novel approach to design of cis-acting DNA structural element for regulation of gene expression in vivo. In: Current Science (Bangalore), 60 (9-10). pp. 586-591.

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Abstract

Taking advantage of the degeneracy of the genetic code we have developed a novel approach to introduce, within a gene, DNA sequences capable of adopting unusual structures and to investigate the role of such sequences in regulation of gene expression in vivo. We used a computer program that generates alternative codon sequences for the same amino-acid sequence to convert a stretch of nucleotides into an inverted-repeat sequence with the potential to adopt cruciform structure. This approach was used to replace a 51-base-pair EcoRI-HindIII segment in the N-terminal region of the beta-galactosidase gene in plasmid pUC19 with a 51-bp synthetic oligonucleotide sequence with the potential to adopt a cruciform structure with 18 bp in the stem region. In selecting the 51-bp sequence, care was taken to include those codons that are preferred in E. coli. E. coli DH5-alpha cells harbouring the plasmid containing the redesigned sequence showed drastic reduction in expression of the beta-galactosidase gene compared to cells harbouring the plasmid with the native sequence. This approach demonstrates the possibility of introducing DNA secondary-structure elements to alter regulation of gene expression in vivo.

Item Type:	Journal Article
Publication:	Current Science (Bangalore)
Publisher:	Indian Academy of Sciences
Additional Information:	Copyright of this article belongs to Indian Academy of Sciences.
Keywords:	Rna Polymerase-Ii;C-Myc Gene;Form-V Dna;Transcriptional Block;Cruciform Dna;Sequences; Binding;Conformation; Termination;Attenuation.
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit Division of Biological Sciences > Molecular Reproduction, Development & Genetics
Date Deposited:	11 Nov 2010 06:44
Last Modified:	11 Nov 2010 06:44
URI:	http://eprints.iisc.ac.in/id/eprint/33627

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