PMID: 9177256Jun 10, 1997Paper

Cloning gibberellin dioxygenase genes from pumpkin endosperm by heterologous expression of enzyme activities in Escherichia coli

Proceedings of the National Academy of Sciences of the United States of America
Theo Lange

Abstract

Gibberellin (GA) plant hormones are biosynthesized via complex pathways, the final steps of which are catalyzed by 2-oxoglutarate-dependent dioxygenases. Here, the cloning of two such enzymes, the GA 7-oxidase and the GA 20-oxidase, is reported using a novel approach, namely, by screening for GA dioxygenase activities expressed as T7 gene 10 fusion proteins in recombinant Escherichia coli. In vitro translation products of mRNA from endosperm of immature pumpkin seeds contained three GA dioxygenase activities, including 7-oxidase, 20-oxidase, and 3beta-hydroxylase. A cDNA expression library was prepared from the endosperm mRNA in lambdaMOSElox. An aliquot of the amplified library was converted to plasmids in vivo and used for transformation of E. coli BL21(DE3), which thereafter expressed recombinant fusion proteins containing 7-oxidase, 20-oxidase, and 3beta-hydroxylase activities. By screening for specific GA dioxygenase expression, clones harboring 7-oxidase and 20-oxidase cDNA were isolated. The ORF of the 7-oxidase cDNA is 945 bp long, encoding for 314 amino acid residues with a calculated Mr of 35,712 and pI of 5.7. Recombinant GA 7-oxidase oxidizes GA12-aldehyde to GA12 and GA14-aldehyde to GA14. Evidence was obtained for...Continue Reading

References

Jan 1, 1990·Methods in Enzymology·L Gold
Sep 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·T Sato, A Theologis
Jun 5, 1983·Journal of Molecular Biology·D Hanahan
Jul 3, 1995·Proceedings of the National Academy of Sciences of the United States of America·Y L XuJ A Zeevaart
Feb 1, 1995·The Plant Cell·H H ChiangH M Goodman
Aug 30, 1994·Proceedings of the National Academy of Sciences of the United States of America·T LangeJ E Graebe
Jun 11, 1993·Nucleic Acids Research·D I Israel
Aug 1, 1996·Current Genetics·V HomannB Tudzynski
Jan 1, 1996·Planta·R Neyland, L E Urbatsch
Jun 1, 1997·Annual Review of Plant Physiology and Plant Molecular Biology·Peter Hedden, Yuji Kamiya
Jun 1, 1996·Annual Review of Plant Physiology and Plant Molecular Biology·Andy G. Prescott, Philip John

❮ Previous
Next ❯

Citations

Aug 23, 2001·Archives of Biochemistry and Biophysics·R LukacinS Martens
Dec 20, 2000·Trends in Plant Science·P Hedden, A L Phillips
Jul 27, 1999·The Plant Journal : for Cell and Molecular Biology·D R LesterJ B Reid
Jan 24, 2012·Journal of Experimental Botany·Maria João Pimenta LangeTheo Lange
Jan 17, 2003·Plant Physiology·Sandra E DavidsonJames B Reid
Mar 20, 2003·Plant Physiology·Andrea FrisseTheo Lange
Aug 30, 2005·Plant Physiology·Theo LangeMaria João Pimenta Lange
Nov 11, 1999·Plant Physiology·D N MartinP Hedden
Apr 26, 2001·Proceedings of the National Academy of Sciences of the United States of America·M C RojasB Tudzynski
Apr 14, 1999·Proceedings of the National Academy of Sciences of the United States of America·S G ThomasP Hedden
Jul 22, 1998·Proceedings of the National Academy of Sciences of the United States of America·C A HelliwellE S Dennis
Mar 20, 2013·Phytochemistry·Maria João Pimenta LangeTheo Lange
Feb 20, 2014·The Plant Journal : for Cell and Molecular Biology·Yosuke KawaiMasaharu Mizutani
Jun 30, 2006·Plant Biology·M J Pimenta Lange, T Lange
Apr 27, 2012·The Biochemical Journal·Peter Hedden, Stephen G Thomas
Oct 28, 2014·Frontiers in Plant Science·Scott C Farrow, Peter J Facchini
Nov 15, 2016·Nature Chemical Biology·Ryan S NettReuben J Peters
Nov 3, 2009·Nature Genetics·Sanwen HuangSonggang Li
Aug 3, 2005·Journal of Experimental Botany·Beata A WoluckaDirk Inzé
Jul 10, 2019·Plant Molecular Biology·Jorge A Condori-ApfataAdriano Nunes-Nesi
Apr 12, 2002·The Journal of Biological Chemistry·Bettina TudzynskiPeter Hedden
Apr 29, 2020·Plant & Cell Physiology·Theo Lange, Maria João Pimenta Lange
Apr 30, 2020·The Journal of Biological Chemistry·Maria João Pimenta LangeTheo Lange

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.