title: The effects of protein solubility on the RNA Integrity Number (RIN) for recombinantEscherichia coli.
abstract: High quality, intact messenger RNA (mRNA) is required for DNA microarray and reverse transcriptase polymerase chain reaction analysis and is generally obtained from total RNA isolations. The most widely recognized measure of RNA integrity is the RNA Integrity Number (RIN) obtained from the Agilent Bioanalyzer, as it provides sizing, quantification, and quality control measures. This work describes comparisons of the RIN values obtained for recombinantE. coli. Uninduced recombinantE. colicultures were examined, as well as induced cultures that produced either a soluble or insoluble recombinant protein. The uninduced cultures and the induced cultures producing soluble protein had higher RIN values than the induced cultures producing insoluble protein. These lower RIN values forE. coliproducing the insoluble protein indicate that cellular degradation of the ribosomal RNA species is the likely cause of the lower RIN values. As the use of DNA microarrays and other gene expression tools increase in usage in the industrial recombinant protein production community, these results suggest the need for further studies to determine acceptable RIN ranges for gene expression analysis and effects of various culture conditions on RIN values for recombinantE. coli.
authors.affiliation: Department of Chemistry, Clemson University.
authors.forename: Mary Alice
authors.initials: MA
authors.lastname: Salazar
authors.affiliation: Department of Chemistry, Clemson University.
authors.forename: Lawrence P
authors.initials: LP
authors.lastname: Fernando
authors.affiliation: Department of Bioengineering, Clemson University.
authors.forename: Faraz
authors.initials: F
authors.lastname: Baig
authors.affiliation: Department of Bioengineering, Clemson University.
authors.forename: Sarah W
authors.initials: SW
authors.lastname: Harcum
keywords: Aggregation; Inclusion bodies; Protease; Purification; RNA; Recombinant DNA