Use of T7 RNA polymerase to direct expression of cloned genes.

A complementary DNA for the Aequorea victoria green fluorescent protein (GFP) produces a fluorescent product when expressed in prokaryotic (Escherichia coli) or eukaryotic (Caenorhabditis elegans) cells. Because exogenous substrates and cofactors are not required for this fluorescence, GFP expression can be used to monitor gene expression and protein localization in living organisms.

http://science.sciencemag.org/content/sci/263/5148/802.full.pdf

Green fluorescent protein as a marker for gene expression

Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes

http://wolfson.huji.ac.il/expression/local/bacteria/studierAutoInduced.pdf

Protein production by auto-induction in high-density shaking cultures

The gene product of the ob locus is important in the regulation of body weight. The ob product was shown to be present as a 16-kilodalton protein in mouse and human plasma but was undetectable in plasma from C57BL/6J ob/ob mice. Plasma levels of this protein were increased in diabetic (db) mice, a mutant thought to be resistant to the effects of ob. Daily intraperitoneal injections of either mouse or human recombinant OB protein reduced the body weight of ob/ob mice by 30 percent after 2 weeks of treatment with no apparent toxicity but had no effect on db/db mice. The protein reduced food intake and increased energy expenditure in ob/ob mice. Injections of wild-type mice twice daily with the mouse protein resulted in a sustained 12 percent weight loss, decreased food intake, and a reduction of body fat from 12.2 to 0.7 percent. These data suggest that the OB protein serves an endocrine function to regulate body fat stores.

https://science.sciencemag.org/content/sci/269/5223/543.full.pdf

Weight-Reducing Effects of the Plasma Protein Encoded by the obese Gene

Vectors for selective expression of cloned DNAs by T7 RNA polymerase

Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements

https://www.cell.com/cell/pdf/S0092-8674(04)01159-6.pdf

Defining the CREB Regulon: A Genome-Wide Analysis of Transcription Factor Regulatory Regions

Ralstonia metallidurans, formerly known as Alcaligenes eutrophus and thereafter as Ralstonia eutropha, is a β-Proteobacterium colonizing industrial sediments, soils or wastes with a high content of heavy metals. The type strain CH34 carries two large plasmids (pMOL28 and pMOL30) bearing a variety of genes for metal resistance. A chronological overview describes the progress made in the knowledge of the plasmid-borne metal resistance mechanisms, the genetics of R. metallidurans CH34 and its taxonomy, and the applications of this strain in the fields of environmental remediation and microbial ecology. Recently, the sequence draft of the genome of R. metallidurans has become available. This allowed a comparison of these preliminary data with the published genome data of the plant pathogen Ralstonia solanacearum, which harbors a megaplasmid (of 2.1 Mb) carrying some metal resistance genes that are similar to those found in R. metallidurans CH34. In addition, a first inventory of metal resistance genes and operons across these two organisms could be made. This inventory, which partly relied on the use of proteomic approaches, revealed the presence of numerous loci not only on the large plasmids pMOL28 and pMOL30 but also on the chromosome. It suggests that metal-resistant Ralstonia, through evolution, are particularly well adapted to the harsh environments typically created by extreme anthropogenic situations or biotopes.

http://dzumenvis.nic.in/Taxonomy/pdf/Ralstonia%20metallidurans,%20a%20bacterium%20specifically.pdf

John J. Dunn

Papers

Use of T7 RNA polymerase to direct expression of cloned genes.

Vectors for selective expression of cloned DNAs by T7 RNA polymerase

Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements

Defining the CREB Regulon: A Genome-Wide Analysis of Transcription Factor Regulatory Regions

Ralstonia metallidurans, a bacterium specifically adapted to toxic metals: towards a catalogue of metal-responsive genes