Vector construction for production of armored nucleic acids

Abstract

A widespread use of RT-PCR as an effective method of RNA-viruses diagnostics has created a need for stable and relevant controls. Unfortunately, controls based on plasmids and vial viruses have several critical disadvantages. The armored RNA technology shows ways to avoid these limitations. Armored RNA is a complex of MS2 bacteriophage coat protein and RNA produced in Escherichia coli by the induction of an expression plasmid that encodes the coat protein and an RNA control sequence. The RNA sequences are protected from RNase digestion within the pseudoviral complex. These particles can work as a control on every stage of PCR virus detection - lysis, purification, revert transcription and amplification. This article describes the process of development and construction of armored RNA for a subsequent use as control for RT-PCR detection of RNA-viruses.