A method for purifying obligate intracellular Coxiella burnetii that employs digitonin lysis of host cells

被引：37

作者：

Cockrell, Diane C. ^{[1
]}

Beare, Paul A. ^{[1
]}

Fischer, Elizabeth R. ^{[2
]}

Howe, Dale ^{[1
]}

Heinzen, Robert. A. ^{[1
]}

机构：

[1] Natl Inst Hlth, Natl Inst Allergy & Infect Dis, Rocky Mt Labs, Intracellular Parasites Lab,Coxiella Pathogenesis, Hamilton, MT 59840 USA

[2] Natl Inst Hlth, Natl Inst Allergy & Infect Dis, Rocky Mt Labs, Res Technol Branch,Microscopy Unit,Res Technol Br, Hamilton, MT 59840 USA

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 2008年 / 72卷 / 03期

关键词：

digitonin; Coxiella; Q fever; cell lysis; purification; vacuole; infectivity; sonication; intracellular parasite;

D O I：

10.1016/j.mimet.2007.12.015

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Purification of the obligate intracellular bacterium Coxiella burnetii requires physical disruption of infected cells. Here we describe a gentle and safe digitonin lysis procedure to release C. burnetii from infected cells. The purity, yield, and infectivity of digitonin-prepped organisms are comparable to that of organisms purified using cell lysis by sonication. Published by Elsevier B.V.

引用

收藏

页码：321 / 325

页数：5

相关论文

共 22 条

[1] ACID-PHOSPHATASE-ACTIVITY IN COXIELLA-BURNETII - A POSSIBLE VIRULENCE FACTOR [J].

BACA, OG ;

ROMAN, MJ ;

GLEW, RH ;

CHRISTNER, RF ;

BUHLER, JE ;

ARAGON, AS .

INFECTION AND IMMUNITY, 1993, 61 (10) :4232-4239

[2] Q-FEVER AND COXIELLA-BURNETII - A MODEL FOR HOST-PARASITE INTERACTIONS [J].

BACA, OG ;

PARETSKY, D .

MICROBIOLOGICAL REVIEWS, 1983, 47 (02) :127-149

[3] Coxiella burnetii localizes in a Rab7-labeled compartment with autophagic characteristics [J].

Berón, W ;

Gutierrez, MG ;

Rabinovitch, M ;

Colombo, MI .

INFECTION AND IMMUNITY, 2002, 70 (10) :5816-5821

[4] Both inducible nitric oxide synthase and NADPH oxidase contribute to the control of virulent phase I Coxiella bumetii infections [J].

Brennan, RE ;

Russell, K ;

Zhang, GQ ;

Samuel, JE .

INFECTION AND IMMUNITY, 2004, 72 (11) :6666-6675

[5] PURIFICATION AND PARTIAL CHARACTERIZATION OF THE MAJOR OUTER-MEMBRANE PROTEIN OF CHLAMYDIA-TRACHOMATIS [J].

CALDWELL, HD ;

KROMHOUT, J ;

SCHACHTER, J .

INFECTION AND IMMUNITY, 1981, 31 (03) :1161-1176

[6] Temporal analysis of Coxiella burnetii morphological differentiation [J].

Coleman, SA ;

Fischer, ER ;

Howe, D ;

Mead, DJ ;

Heinzen, RA .

JOURNAL OF BACTERIOLOGY, 2004, 186 (21) :7344-7352

[7] Proteome and antigen profiling of Coxiella burnetii developmental forms [J].

Coleman, Sherry A. ;

Fischer, Elizabeth R. ;

Cockrell, Diane C. ;

Voth, Daniel E. ;

Howe, Dale ;

Mead, David J. ;

Samuel, James E. ;

Heinzen, Robert A. .

INFECTION AND IMMUNITY, 2007, 75 (01) :290-298

[8] Rickettsia rickettsii infection of the EA.hy 926 endothelial cell line:: morphological response to infection and evidence for oxidative injury [J].

Eremeeva, ME ;

Silverman, DJ .

MICROBIOLOGY-SGM, 1998, 144 :2037-2048

[9] Coxiella burnetii inhabits a cholesterol-rich vacuole and influences cellular cholesterol metabolism [J].

Howe, D ;

Heinzen, RA .

CELLULAR MICROBIOLOGY, 2006, 8 (03) :496-507

[10] LABORATORY-ACQUIRED Q FEVER - A REPORT OF 50 CASES [J].

JOHNSON, JE ;

KADULL, PJ .

AMERICAN JOURNAL OF MEDICINE, 1966, 41 (03) :391-+