Crude protein extraction protocol for phage N15 protelomerase in vitro enzymatic assays

被引：5
|
作者：
Chen, Qingwen [1 ]
Narayanan, Kumaran [1 ,2 ]
机构：
[1] Monash Univ, Sch Sci, Bandar Sunway 46150, Malaysia
[2] Mt Sinai Sch Med, Dept Genet & Genom Sci, New York, NY USA
来源：
ANALYTICAL BIOCHEMISTRY | 2011年 / 414卷 / 01期
关键词：
AGROBACTERIUM-TUMEFACIENS C58; LINEAR PLASMID PROPHAGE; COVALENTLY CLOSED ENDS; BORRELIA-BURGDORFERI; ESCHERICHIA-COLI; GENOMIC SEQUENCE; REPLICATION; EXCHANGE; SYSTEM; SITE;
D O I：
10.1016/j.ab.2011.03.006
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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. (C) 2011 Elsevier Inc. All rights reserved.
引用
收藏
页码：169 / 171
页数：3
相关论文
共 50 条
  • [31] Investigating the effects of sex of growing Nellore cattle and crude protein intake on the utilization of recycled N for microbial protein synthesis in the rumen by using intravenous 15N15N-urea infusion
    Prates, L. L.
    Valadares, R. F. D.
    Valadares Filho, S. C.
    Detmann, E.
    Ouellet, D. R.
    Batista, E. D.
    Zanetti, D.
    Pacheco, M. V. C.
    Silva, B. C.
    ANIMAL FEED SCIENCE AND TECHNOLOGY, 2017, 231 : 119 - 130
  • [32] Dry matter, crude protein and gross energy digestibility of corn silage and concentrate mixture through in vivo and in vitro digestion assays
    de Oliveira, MD
    Sampaio, AAM
    Vieira, PD
    Banzatto, DA
    REVISTA DA SOCIEDADE BRASILEIRA DE ZOOTECNIA-JOURNAL OF THE BRAZILIAN SOCIETY OF ANIMAL SCIENCE, 1997, 26 (03): : 590 - 594
  • [33] Protocol for the upgraded high-throughput SPME system for biocompatible in vitro extraction from small volume for metabolomics and pharmaceutical assays
    Szeliska, Paulina
    Jaroch, Karol
    Charemski, Bartlomiej
    Kahremanoglu, Kubra
    Cretin, Enes
    Boyaci, Ezel
    Bojko, Barbara
    GREEN ANALYTICAL CHEMISTRY, 2025, 13
  • [34] Modeling and Optimizing a New Culture Medium for In Vitro Rooting of G×N15 Prunus Rootstock using Artificial Neural Network-Genetic Algorithm
    Mohammad Mehdi Arab
    Abbas Yadollahi
    Maliheh Eftekhari
    Hamed Ahmadi
    Mohammad Akbari
    Saadat Sarikhani Khorami
    Scientific Reports, 8
  • [35] Predicting omasal flow of nonammonia N and milk protein yield from in vitro-determined utilizable crude protein at the duodenum
    Gidlund, H.
    Vaga, M.
    Ahvenjarvi, S.
    Rinne, M.
    Ramin, M.
    Huhtanen, P.
    JOURNAL OF DAIRY SCIENCE, 2018, 101 (02) : 1164 - 1176
  • [36] Extraction of Crude Bromelain from Pineapple (Ananas comosus L.) Fruit Waste and its in vitro Protein Digestibility
    Wiboonsiriku, J.
    Khuwijitjaru, P.
    Klahan, R.
    JOURNAL OF AGRICULTURAL SCIENCES, 2024, 19 (01): : 73 - 88
  • [37] Modeling and Optimizing a New Culture Medium for In Vitro Rooting of G X N15 Prunus Rootstock using Artificial Neural Network-Genetic Algorithm
    Arab, Mohammad Mehdi
    Yadollahi, Abbas
    Eftekhari, Maliheh
    Ahmadi, Hamed
    Akbari, Mohammad
    Khorami, Saadat Sarikhani
    SCIENTIFIC REPORTS, 2018, 8
  • [38] The use of 15N-labelled protein to account for the endogenous nitrogen contribution to in vitro protein digestibility measurement
    Menard, Olivia
    Chauvet, Lucile
    Henry, Gwenaele
    Dupont, Didier
    Gaudichon, Claire
    Calvez, Juliane
    Deglaire, Amelie
    FOOD RESEARCH INTERNATIONAL, 2023, 173
  • [39] Use of central composite design to optimize working conditions of Streptomyces griseus enzymatic method in estimating in vitro rumen undegraded crude protein of feedstuffs
    Gallo, A.
    Fortunati, P.
    Bruschi, S.
    Giuberti, G.
    Masoero, F.
    JOURNAL OF AGRICULTURAL SCIENCE, 2018, 156 (01): : 100 - 109
  • [40] DYNAMICS OF THE AMINO-ACID AND PROTEIN-METABOLISM OF LAYING HENS AFTER THE APPLICATION OF N-15 LABELED WHEAT-PROTEIN .10. N-15 LABELING OF THE CRUDE PROTEIN AND THE AMINO-ACIDS OF THE OVIDUCT
    GRUHN, K
    HENNIG, A
    ARCHIV FUR TIERERNAHRUNG-ARCHIVES OF ANIMAL NUTRITION, 1989, 39 (03): : 311 - 320
12345