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Proc. Natl. Acad. Sci. USA 103, 2827–32. The U(L)41 protein of herpes simplex virus 1 degrades RNA by endonucleolytic cleavage in absence of other cellular or viral proteins. 2006

Taddeo, B., Zhang, W. and Roizman, B.

Notes: In this study, an in vitro assay was performed to examine the ribonuclease activity of the herpes simplex virus virion host shutoff (vhs) protein. To test whether a purified GST-vhs fusion protein exhibited RNase activity, the entire 3’ UTR of human IEX-1 mRNA (a known target of vhs) was labeled with 32P and then incubated at 30°C with either GST-vhs or GST alone. Samples were taken at various time points and analyzed by denaturing polyacrylamide gel electrophoresis and autoradiography. The GST-vhs fusion protein displayed RNase activity. When RNasin® Plus RNase Inhibitor was added to the reaction, no degradation products were seen, confirming that the nuclease activity of GST-vhs could be blocked by an RNase inhibitor. (3391)

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Clin. Exp. Immunol. 131, 144–51. Contribution of Vα24+Vβ11+ natural killer T cells in Wilsonian hepatitis. 2005

Kinebuchi, M., Matsuura, A., Ohya, K., Abo, W. and Kitazawa, J.

Notes: To prepare single rat NK cells for reverse transcription, the cells were sorted by flow cytometry and lysed using 0.5% NP-40, 2.2µl of 5X M-MLV Reverse Transcriptase Buffer, 0.3µl of 0.1mol/l DTT, 5.5µl DEPC-treated water and 1U of RNasin® Plus RNase Inhibitor. After a 30-minute incubation on ice, the lysates were heated to 65°C for 90 seconds, cooled to 22°C for 3 minutes and placed back on ice. For the RT reaction, 1U RNasin® Plus RNase Inhibitor, 60 U of M-MLV Reverse Transcriptase, 2.5mmol/l dNTPs and 100µM primers were added to these single-cell lysates. (3390)

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Mol. Cell. Biol. 25, 621–636. Mutations in the RNA polymerase III subunit Rpc11p that decrease RNA 3’ cleavage activity increase 3’-terminal oligo(U) length and La-dependent tRNA processing. 2005

Huang, Y., Intine, R.V., Mozlin, A., Hasson, S. and Maraia, R.J.

Notes: RNasin® Plus RNase Inhibitor was added to RNA-DNA ligation reactions and in vitro transcription reactions to protect RNA molecules in both reactions.  For in vitro transcription reactions, 1 unit of RNasin® Plus RNase Inhibitor was added to the reaction buffer.   (3257)

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