Academic journal article Iranian Journal of Public Health

Design and Cloning of an Antisense against Bacteriorhodopsin-Linked Protein Coding Gene of Halobacterium Salinarum R1

Academic journal article Iranian Journal of Public Health

Design and Cloning of an Antisense against Bacteriorhodopsin-Linked Protein Coding Gene of Halobacterium Salinarum R1

Article excerpt

Background: Our goal in this study is designing antisense against blp . To identify function of genes there are multiple ways. One of these approaches is designing of antisense that prevent function of desired gene.

Methods: At the first stage we designed a primer for a 180 bp area that contains ribosome binding site and initial part of gene. Then restriction sites of BamH1 and Xba1 inserted at the 5' of them with 5 random nucleotides at the end of each primers. The primer was analyzed be oligoanalyzer software. We amplify 180 bp fragment by PCR and cloned it in pBluscript SKII cloning vector. Then 180 bp subcloned into shuttle vector pVDSH1, that is a vector contains promoter and terminator sequences and our 180 bp fragment between these essential elements.

Results: The results of digestion and sequencing showed that this 180 bp fragment was inserted into vector in inverse direction and placed downstream of an inducible promoter and with manipulating of KCL concentration, we can control the rate of final target protein production and consequently evaluate the rate of silencing of the gene. …

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