Increasing Expression of an Anti-Picloram Single-Chain Variable Fragment (ScFv) Antibody and Resistance to Picloram in Transgenic Tobacco (<i>Nicotiana tabacum</i>)

SummaryThe phytohormone auxin has been known for >50 years to be required for entry into the cell cycle. Despite the critical effects exerted by auxin on the control of cell division, the molecular mechanism by which auxin controls this pathway is poorly understood, and how auxin is perceived upstream of any change in the cell cycle is unknown. Auxin Binding Protein 1 (ABP1) is considered to be a candidate auxin receptor, triggering early modification of ion fluxes across the plasma membrane in response to auxin. ABP1 has also been proposed to mediate auxin-dependent cell expansion, and is essential for early embryonic development. We investigated whether ABP1 has a role in the cell cycle. Functional inactivation of ABP1 in the model plant cell system BY2 was achieved through cellular immunization via the conditional expression of a single-chain fragment variable (scFv). This scFv was derived from a well characterized anti-ABP1 monoclonal antibody previously shown to block the activity of the protein. We demonstrate that functional inactivation of ABP1 results in cell-cycle arrest, and provide evidence that ABP1 plays a critical role in regulation of the cell cycle by acting at both the G1/S and G2/M checkpoints. We conclude that ABP1 is essential for the auxin control of cell division and is likely to constitute the first step of the auxin-signalling pathway mediating auxin effects on the cell cycle.

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“…, 2005) or to confer pathogen or herbicide resistance (Eto et al. , 2003; Olea‐Popelka et al. , 2005; Villani et al.…”

Section: Introductionmentioning

confidence: 99%

The auxin‐binding protein 1 is essential for the control of cell cycle

et al. 2007

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“…The V H and V L sequences of the S20 IgG2 (22), i.e., the first 119 and 113 amino acids of GenBank accession numbers AAK39434 and AAK39435, respectively, were combined in silico with the gamma-1 and kappa constantregion sequences, i.e., the entire 330 and 106 amino acids of GenBank accession numbers P01857 (18) and P01834 (20), respectively. Complete coding sequences for both HC and LC polypeptides were optimized for expression in transgenic tobacco and synthesized according to the method of Olea-Popelka et al (36) and Almquist et al (1). The gamma-1 sequence had six-His and KDEL ER retrieval signal (43) motifs added to its carboxyl terminus.…”

Section: Methodsmentioning

confidence: 99%

A Human Anti-Pseudomonas aeruginosaSerotype O6ad Immunoglobulin G1 Expressed in Transgenic Tobacco Is Capable of Recruiting Immune System Effector Function In Vitro

McLean

Almquist

Niu

et al. 2007

Antimicrob Agents Chemother

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The production of a recombinant human IgG1 in transgenic tobacco was examined to determine whether a plant-derived antibody could recruit immune system effector function against a bacterial pathogen. A plant transformation vector was engineered to contain genes for a human kappa light chain and a human gamma-1 heavy chain with V H and V L sequences from a previously identified human IgG2 monoclonal antibody (MAb) that specifically binds to and opsonizes Pseudomonas aeruginosa serotype O6ad. Unique NcoI and NotI restriction sites were incorporated to flank these variable sequences, resulting in a plant transformation vector that could be engineered for expression of any other human IgG1 antibody, requiring only the substitution of other V H and V L antigen-binding coding sequences. The plant-produced IgG1 was determined to have highmannose glycan content and to be capable of mediating opsonophagocytosis of P. aeruginosa serotype O6ad in vitro using human complement and human polymorphonuclear leukocytes. Thus, MAbs produced in plants from this vector could provide human IgG1 MAbs for targeting other pathogens that require the recruitment of immune system effector functions.

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“…In addition, plants and crop species have the potential to enable extremely cost effective and efficient production of Abs [22].…”

Section: Immunorecognitionmentioning

confidence: 99%

Chemiluminescence immunoassay

Zhao

Sun

Chu

2009

TrAC Trends in Analytical Chemistry

186

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Chemiluminescence immunoassay (CLIA) offers several potential advantages and has applications in clinical chemistry, bioanalysis and environmental analysis. This review covers recent developments and applications of CLIA in immunorecognition, CL markers and related techniques, and solid-phase materials. We describe automation, integration and miniaturization of CLIA. We evaluate different CLIA systems and give some pointers as to the likely directions of future developments. ª

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Increasing Expression of an Anti-Picloram Single-Chain Variable Fragment (ScFv) Antibody and Resistance to Picloram in Transgenic Tobacco (Nicotiana tabacum)

Cited by 22 publications

References 31 publications

The auxin‐binding protein 1 is essential for the control of cell cycle

The auxin‐binding protein 1 is essential for the control of cell cycle

A Human Anti-Pseudomonas aeruginosaSerotype O6ad Immunoglobulin G1 Expressed in Transgenic Tobacco Is Capable of Recruiting Immune System Effector Function In Vitro

Chemiluminescence immunoassay

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