TY - JOUR
T1 - Chapter 6
T2 - Mycobacterium: Isolation, Maintenance, Transformation, and Mutant Selection
AU - Connell, Nancy D.
PY - 1995/1/1
Y1 - 1995/1/1
N2 - This chapter discusses methods for culturing Mycobacterium, isolating genetic mutants, and moving DNA in and out of the cells. Experimental methods for culturing and genetically manipulating M. avium, M. tuberculosis, M. smegmatis, and bacille Calmette Guerin (BCG) is discussed in the chapter. The isolation of mycobacteria from contaminated sources requires decontamination because specimens usually contain mixed bacterial flora and some degree of organic debris such as body fluids, blood cells, and tissue fragments. There are a number of isolation protocols but all require digestion, decontamination, and concentration steps. The mycobacteria must be recovered undamaged while organic debris is liquefied and contaminating micororganisms are killed. The chapter discusses protocol for treatment of sputum samples and is suitable for use in clinical biosafety level (BSL-2) and experimental BSL-3 laboratories for the treatment of clinical samples. The mycobacteriophages have been isolated and developed for the purpose of strain typing. Their use in transfer of genetic material for the construction of strains is discussed.
AB - This chapter discusses methods for culturing Mycobacterium, isolating genetic mutants, and moving DNA in and out of the cells. Experimental methods for culturing and genetically manipulating M. avium, M. tuberculosis, M. smegmatis, and bacille Calmette Guerin (BCG) is discussed in the chapter. The isolation of mycobacteria from contaminated sources requires decontamination because specimens usually contain mixed bacterial flora and some degree of organic debris such as body fluids, blood cells, and tissue fragments. There are a number of isolation protocols but all require digestion, decontamination, and concentration steps. The mycobacteria must be recovered undamaged while organic debris is liquefied and contaminating micororganisms are killed. The chapter discusses protocol for treatment of sputum samples and is suitable for use in clinical biosafety level (BSL-2) and experimental BSL-3 laboratories for the treatment of clinical samples. The mycobacteriophages have been isolated and developed for the purpose of strain typing. Their use in transfer of genetic material for the construction of strains is discussed.
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U2 - 10.1016/S0091-679X(08)61848-8
DO - 10.1016/S0091-679X(08)61848-8
M3 - Article
C2 - 7707982
AN - SCOPUS:0028693996
SN - 0091-679X
VL - 45
SP - 107
EP - 125
JO - Methods in cell biology
JF - Methods in cell biology
IS - C
ER -