Recombinant Gene Expression: Reviews and Protocols

Front Cover
Paulina Balbas, Argelia Lorence
Springer Science & Business Media, Feb 4, 2008 - Science - 508 pages
Since newly created beings are often perceived as either wholly good or bad, the genetic alteration of living cells impacts directly on a symbolic meaning deeply imbedded in every culture. During the earlier years of gene expression research, te- nological applications were confined mainly to academic and industrial laboratories, and were perceived as highly beneficial since molecules that were previously unable to be separated or synthesized became accessible as therapeutic agents. Such were the success stories of hormones, antibodies, and vaccines produced in the bacterium Escherichia coli. Originally this bacterium gained fame among humans for being an unwanted host in the intestine, or worse yet, for being occasionally dangerous and pathogenic. H- ever, it was easily identified in contaminated waters during the 19th century, thus becoming a clear indicator of water pollution by human feces. Tamed, cultivated, and easily maintained in laboratories, its fast growth rate and metabolic capacity to adjust to changing environments fascinated the minds of scientists who studied and modeled such complex phenomena as growth, evolution, genetic exchange, infection, survival, adaptation, and further on—gene expression. Although at the lower end of the complexity scale, this microbe became a very successful model system and a key player in the fantastic revolution kindled by the birth of recombinant DNA technology.
 

Contents

Challenges and Solutions
15
Chromosomal Chromosomal
37
FoldingPromoting Agents in Recombinant Protein Production
53
Transformation
55
pBR322 and Protein Expression Systems in E coli
77
αComplementation Enabled T7 Expression Vectors
91
7
113
13
183
Engineering the Chloroplast Genome for Hyperexpression
365
Group
372
New Selection Marker for Plant Transformation
385
Enhancer Detection and Gene Trapping as Tools
397
General editing
405
Gene Transfer and Expression in Mammalian Cell Lines
417
chromosomal
425
Sustained Heterologous Transgene Expression in Mammalian
435

Gene Transfer and Expression of Recombinant Proteins
209
16
241
Controlled Expression of Homologous Genes
259
HighThroughput Expression in Microplate Format
267
HighThroughput Expression in Microplate Format
277
Multiple Gene Expression by Chromosomal Integration
287
Novel Technologies
315
Gene Transfer and Expression in Plants
329
in Plastic Sleeve Bioreactors
351
Selection
438
Inducible Gene Expression in Mammalian Cells and Mice
451
FlpMediated Integration of Expression Cassettes into FRTTagged
467
Fungi
472
Generation of HighRecombinant ProteinProducing
477
Preparation of Recombinant Proteins in Milk
485
2022
486
Index
495
Copyright

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