A RAPID ELECTROPHORETIC METHOD FOR THE MEASUREMENT OF PLASMID COPY NUMBER

被引：19

作者：

LEWINGTON, J

DAY, MJ

机构：

[1] Department of Applied Biology, University of Wales, Institute of Science and Technology, PO Box 13, Cathays Park, Cardiff,CF1 3XF, United Kingdom

来源：

LETTERS IN APPLIED MICROBIOLOGY | 1986年 / 3卷 / 06期

关键词：

D O I：

10.1111/j.1472-765X.1986.tb01561.x

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A rapid and inexpensive method for the measurement of copy number of small plasmids, ranging from 8-7 to 13 kb, in under 1 ml of liquid culture is described. The method involves whole cell lysis, electrophoretic separation of plasmid and chromosomal DNA followed by relative densitometric measurement of each, to give an estimation of the plasmid copies per chromosome. Results can be obtained in under 8 h and the method proved to be reproducible, fast and ideal for processing large numbers of samples from batch or continuous culture. © 1986 Oxford University Press. All rights reserved.

引用

页码：109 / 112

页数：4

共 9 条

[1] BACTERIAL GENOME SIZES DETERMINED BY DNA RENATURATION STUDIES [J].

BAK, AL ;

CHRISTIANSEN, C ;

STENDERUP, A .

JOURNAL OF GENERAL MICROBIOLOGY, 1970, 64 (DEC) :377-+

[2] COMPARISON OF DEOXYRIBONUCLEIC-ACID MOLECULAR-WEIGHTS AND HOMOLOGIES OF PLASMIDS CONFERRING LINKED RESISTANCE TO STREPTOMYCIN AND SULFONAMIDES [J].

BARTH, PT ;

GRINTER, NJ .

JOURNAL OF BACTERIOLOGY, 1974, 120 (02) :618-630

[3] INTERACTION OF CLOSED CIRCULAR DNA WITH INTERCALATIVE DYES .I. SUPERHELIX DENSITY OF SV40 DNA IN PRESENCE AND ABSENCE OF DYE [J].

BAUER, W ;

VINOGRAD, J .

JOURNAL OF MOLECULAR BIOLOGY, 1968, 33 (01) :141-&

[4] CIRCULAR DNA FORMS OF COLICINOGENIC FACTORS E1 E2 AND E3 FROM ESCHERICHIA COLI [J].

BAZARAL, M ;

HELINSKI, DR .

JOURNAL OF MOLECULAR BIOLOGY, 1968, 36 (02) :185-&

[5] GENETICS OF PSEUDOMONAS [J].

HOLLOWAY, BW .

BACTERIOLOGICAL REVIEWS, 1969, 33 (03) :419-+

[6] DETERMINATION OF PLASMID COPY NUMBER BY FLUORESCENCE DENSITOMETRY [J].

PROJAN, SJ ;

CARLETON, S ;

NOVICK, RP .

PLASMID, 1983, 9 (02) :182-190

[7]

Shepard H M, 1979, Methods Enzymol, V68, P503

[8] PLASMID COPY NUMBER CONTROL - ISOLATION AND CHARACTERIZATION OF HIGH-COPY-NUMBER MUTANTS OF PLASMID PE194 [J].

WEISBLUM, B ;

GRAHAM, MY ;

GRYCZAN, T ;

DUBNAU, D .

JOURNAL OF BACTERIOLOGY, 1979, 137 (01) :635-643

[9]

WHEATCROFT R, 1981, J GENERAL MICROBIOLO, V124

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