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Full-Text Articles in Molecular Biology
Identification And Expression Analysis Of Methyl Jasmonate Responsive Ests In Paclitaxel Producing Taxus Cuspidata Suspension Culture Cells, Sangram K. Lenka, Nadia Boutaoui, Bibin Paulose, Kham Vongpaseuth, Jennifer Normanly, Susan C. Roberts, Elsbeth L. Walker
Identification And Expression Analysis Of Methyl Jasmonate Responsive Ests In Paclitaxel Producing Taxus Cuspidata Suspension Culture Cells, Sangram K. Lenka, Nadia Boutaoui, Bibin Paulose, Kham Vongpaseuth, Jennifer Normanly, Susan C. Roberts, Elsbeth L. Walker
Jennifer Normanly
Background Taxol® (paclitaxel) promotes microtubule assembly and stabilization and therefore is a potent chemotherapeutic agent against wide range of cancers. Methyl jasmonate (MJ) elicited Taxus cell cultures provide a sustainable option to meet the growing market demand for paclitaxel. Despite its increasing pharmaceutical importance, the molecular genetics of paclitaxel biosynthesis is not fully elucidated. This study focuses on identification of MJ responsive transcripts in cultured Taxus cells using PCR-based suppression subtractive hybridization (SSH) to identify genes involved in global pathway control. Results Six separate SSH cDNA libraries of paclitaxel-accumulating Taxus cuspidata P991 cell lines were constructed at three different post-elicitation …
Contribution Of Taxane Biosynthetic Pathway Gene Expression To Observed Variability In Paclitaxel Accumulation In Taxus Suspension Cultures, Rohan A. Patil, Martin E. Kolewe, Jennifer Normanly, Elsbeth L. Walker, Susan C. Roberts
Contribution Of Taxane Biosynthetic Pathway Gene Expression To Observed Variability In Paclitaxel Accumulation In Taxus Suspension Cultures, Rohan A. Patil, Martin E. Kolewe, Jennifer Normanly, Elsbeth L. Walker, Susan C. Roberts
Jennifer Normanly
Variability in product accumulation is one of the major obstacles limiting the widespread commercialization of plant cell culture technology to supply natural product pharmaceuticals. Despite extensive process engineering efforts, which have led to increased yields, plant cells exhibit variability in productivity that is poorly understood. Elicitation of Taxus cultures with methyl jasmonate (MeJA) induces paclitaxel accumulation, but to varying extents in different cultures. In this work, cultures with different aggregation profiles were established to create predictable differences in paclitaxel accumulation upon MeJA elicitation. Expression of known paclitaxel biosynthetic genes in MeJA-elicited cultures exhibiting both substantial (15-fold) and moderate (2-fold) differences …
The Arabidopsis Atr1 Myb Transcription Factor Controls Indolic Glucosinolate Homeostasis, John L. Celenza, Juan A. Quiel, Gromoslaw A. Smolen, Houra Merrikh, Angela R. Silvestro, Jennifer Normanly, Judith Bender
The Arabidopsis Atr1 Myb Transcription Factor Controls Indolic Glucosinolate Homeostasis, John L. Celenza, Juan A. Quiel, Gromoslaw A. Smolen, Houra Merrikh, Angela R. Silvestro, Jennifer Normanly, Judith Bender
Jennifer Normanly
Plants derive a number of important secondary metabolites from the amino acid tryptophan (Trp), including the growth regulator indole-3-acetic acid (IAA) and defense compounds against pathogens and herbivores. In previous work, we found that a dominant overexpression allele of the Arabidopsis (Arabidopsis thaliana) Myb transcription factor ATR1, atr1D, activates expression of a Trp synthesis gene as well as the Trp-metabolizing genes CYP79B2, CYP79B3, and CYP83B1, which encode enzymes implicated in production of IAA and indolic glucosinolate (IG) antiherbivore compounds. Here, we show that ATR1 over expression confers elevated levels of IAA and IGs. In addition, we show that an atr1 …
Trp-Dependent Auxin Biosynthesis In Arabidopsis: Involvement Of Cytochrome P450s Cyp79b2 And Cyp79b3, Yunde Zhao, Anna K. Hull, Neeru R. Gupta, Kendrick A. Goss, José Alonso, Joseph R. Ecker, Jennifer Normanly, Joanne Chory, John L. Celenza
Trp-Dependent Auxin Biosynthesis In Arabidopsis: Involvement Of Cytochrome P450s Cyp79b2 And Cyp79b3, Yunde Zhao, Anna K. Hull, Neeru R. Gupta, Kendrick A. Goss, José Alonso, Joseph R. Ecker, Jennifer Normanly, Joanne Chory, John L. Celenza
Jennifer Normanly
The plant hormone auxin regulates many aspects of plant growth and development. Although several auxin biosynthetic pathways have been proposed, none of these pathways has been precisely defined at the molecular level. Here we provide in planta evidence that the two Arabidopsis cytochrome P450s, CYP79B2 and CYP79B3, which convert tryptophan (Trp) to indole-3-acetaldoxime (IAOx) in vitro, are critical enzymes in auxin biosynthesis in vivo. IAOx is thus implicated as an important intermediate in auxin biosynthesis. Plants overexpressing CYP79B2 contain elevated levels of free auxin and display auxin overproduction phenotypes. Conversely, cyp79B2 cyp79B3 double mutants have reduced levels of IAA and …