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CPU; AMD Athlon II X2 220 2.8GHz – Dual Core – Cache 1Mo – Bus. AMD Athlon II X3 440 Processor – Driver Download * Vendor: * Product: * B08:01:1D.ization line. The 1^st^-order chemiluminescence measurement was calibrated to the counts per minute for a 2 ng/mL nitrite standard (50 mM NaOH, Sigma) used to obtain a nitrite standard curve (0, 1, 2, 4, 8, 16, 32 ng/mL). Nitrite values were calculated as pg/mL using the nitrite standard curve and by comparison to a standard nitrite solution.
Zymography {#s4_4}
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Conditioned media from *L. naganoensis* cultures was collected and treated with SDS-PAGE sample buffer, and subsequently analysed by electrophoresis on 9% polyacrylamide gels containing 1% gelatin. Gels were washed and incubated with reaction buffer containing 100 mM Tris-HCl (pH 8.0), 200 mM NaCl, 10 mM CaCl~2~, 0.02% NaN~3~, and 1 μg/mL antipain overnight at 37°C. Gels were stained with 0.5% Coomassie blue.
Isolation of *L. naganoensis* RNase P subunit 3 {#s4_5}
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*L. naganoensis* RNase P subunit 3 was isolated by a native gradient in an M-280 FPLC system (Pharmacia) using an Äkta Purifier system (Pharmacia) as described previously \[[@R48]\].
*In vitro* translation {#s4_6}
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*In vitro* translation assays were carried out using the TNT® Quick Coupled Transcription/Translation System according to the manufacturer’s instruction (Promega). Plasmids pUC-rnlA and pUC-rnlA-wt-nuclease were linearized by *Bam*HI. Both plasmids were *in vitro* transcribed using T7 RNA polymerase (Promega) and transcribed RNA was treated with DNase I (Promega). The resulting mRNAs were translated for 90 min at 30°C using TNT® system. The resulting proteins
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