TY - JOUR
T1 - Crude protein extraction protocol for phage N15 protelomerase in vitro enzymatic assays
AU - Chen, Qingwen
AU - Narayanan, Kumaran
PY - 2011
Y1 - 2011
N2 - The phage N15 protelomerase enzyme (TelN) is essential for the replication of its genome by resolution of its telRL domain, located within a telomerase occupancy site (tos), into hairpin telomeres. Isolation of TeIN for in vitro processing of tos, however, is a highly complex process, requiring multiple purification steps. In this study a simplified protocol for crude total protein extraction is described that retains the tos-cleaving activity of TeIN for at least 4 weeks, greatly simplifying in vitro testing of its activity. This protocol may be extended for functional analysis of other phage and bacterial proteins, particularly DNA-processing enzymes
AB - The phage N15 protelomerase enzyme (TelN) is essential for the replication of its genome by resolution of its telRL domain, located within a telomerase occupancy site (tos), into hairpin telomeres. Isolation of TeIN for in vitro processing of tos, however, is a highly complex process, requiring multiple purification steps. In this study a simplified protocol for crude total protein extraction is described that retains the tos-cleaving activity of TeIN for at least 4 weeks, greatly simplifying in vitro testing of its activity. This protocol may be extended for functional analysis of other phage and bacterial proteins, particularly DNA-processing enzymes
UR - http://www.sciencedirect.com/science/article/pii/S0003269711001631
U2 - 10.1016/j.ab.2011.03.006
DO - 10.1016/j.ab.2011.03.006
M3 - Article
SN - 0003-2697
VL - 414
SP - 169
EP - 171
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -