Recombinant Gene Expression: Reviews and ProtocolsPaulina Balbas, Argelia Lorence 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 |
Other editions - View all
Recombinant Gene Expression: Reviews and Protocols Paulina Balbas,Argelia Lorence No preview available - 2010 |
Recombinant Gene Expression: Reviews and Protocols Paulina Balbas,Argelia Lorence No preview available - 2004 |
Common terms and phrases
acid activity Agrobacterium amplification antibiotic antibody Arabidopsis bacteria Balbás Biol Biotechnol buffer cassette cDNA cell lines centrifugation chloroplast chromatin chromosomal cloning concentration containing copy number culture Curr dilution efficiency encoding enhancer Escherichia coli ethanol eukaryotic expression systems expression vector extract folding function fungi fusion gene expression gene transfer gene trap genetic genome growth Halomonadaceae Halomonas halophilic homologous Incubate induced insertion integration lacZ mammalian cells marker gene medium membrane method Microbiol moderately halophilic mutagenesis mutants NaCl Natl pastoris pBAC/oriV PCR product periplasmic plasmid plates polymerase primers promoter protein expression protocol purification reaction recombinant protein recombinant protein production region replication reporter gene restriction enzyme Resuspend SEAP selectable marker sequence soluble solution specific stable sterile strains strategies Subheading supernatant T-DNA target temperature tion tissue transcription transfection transformation transgene expression transgenic plants transposon tube vitro vivo yeast zeocin µg/mL