The art and design of genetic screens:: Escherichia coli

被引:60
作者
Shuman, HA
Silhavy, TJ [1 ]
机构
[1] Princeton Univ, Dept Mol Biol, Princeton, NJ 08544 USA
[2] Columbia Univ, Coll Phys & Surg, Dept Microbiol, New York, NY 10032 USA
基金
美国国家科学基金会;
关键词
D O I
10.1038/nrg1087
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
This article summarizes the general principles of selections and screens in Escherichia coli. The focus is on the lac operon, owing to its inherent simplicity and versatility. Examples of different strategies for mutagenesis and mutant discovery are described. In particular, the usefulness and effectiveness of simple colour-based screens are illustrated. The power of lac genetics can be applied to almost any bacterial system with gene fusions that hook any gene of interest to lacZ, which is the structural gene that encodes beta-galactosidase. The diversity of biological processes that can be studied with lac genetics is remarkable and includes DNA metabolism, gene regulation and signal transduction, protein localization and folding, and even electron transport.
引用
收藏
页码:419 / 431
页数:13
相关论文
共 52 条
[1]   Systematic identification of essential genes by in vitro mariner mutagenesis [J].
Akerley, BJ ;
Rubin, EJ ;
Camilli, A ;
Lampe, DJ ;
Robertson, HM ;
Mekalanos, JJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (15) :8927-8932
[2]   IDENTIFICATION OF A PROTEIN REQUIRED FOR DISULFIDE BOND FORMATION INVIVO [J].
BARDWELL, JCA ;
MCGOVERN, K ;
BECKWITH, J .
CELL, 1991, 67 (03) :581-589
[3]  
BASSFORD P, 1979, NATURE, V277, P538, DOI 10.1038/277538a0
[4]   Genetic control of biochemical reactions in neurospora [J].
Beadle, GW ;
Tatum, EL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1941, 27 :499-506
[5]   Regulation of RpoS proteolysis in Escherichia coli:: The response regulator RssB is a recognition factor that interacts with the turnover element in RpoS [J].
Becker, G ;
Klauck, E ;
Hengge-Aronis, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (11) :6439-6444
[6]  
BECKWITH J, 1963, BIOCHIM BIOPHYS ACTA, V76, P162
[7]   GENETIC-EVIDENCE THAT TN10 TRANSPOSES BY A NON-REPLICATIVE MECHANISM [J].
BENDER, J ;
KLECKNER, N .
CELL, 1986, 45 (06) :801-815
[8]   The complete genome sequence of Escherichia coli K-12 [J].
Blattner, FR ;
Plunkett, G ;
Bloch, CA ;
Perna, NT ;
Burland, V ;
Riley, M ;
ColladoVides, J ;
Glasner, JD ;
Rode, CK ;
Mayhew, GF ;
Gregor, J ;
Davis, NW ;
Kirkpatrick, HA ;
Goeden, MA ;
Rose, DJ ;
Mau, B ;
Shao, Y .
SCIENCE, 1997, 277 (5331) :1453-+
[9]   INTRACELLULAR PROTEIN TOPOGENESIS [J].
BLOBEL, G .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (03) :1496-1500
[10]   TRANSFER OF PROTEINS ACROSS MEMBRANES .1. PRESENCE OF PROTEOLYTICALLY PROCESSED AND UNPROCESSED NASCENT IMMUNOGLOBULIN LIGHT-CHAINS ON MEMBRANE-BOUND RIBOSOMES OF MURINE MYELOMA [J].
BLOBEL, G ;
DOBBERSTEIN, B .
JOURNAL OF CELL BIOLOGY, 1975, 67 (03) :835-851