Transgenic functional complementation assay (Brassicaceae)

Knowledge graph centered on Transgenic functional complementation assay (Brassicaceae) with 24 nodes and 95 connections. Top connected: Ligusticum porteri, Arabidopsis, Boechera stricta, Boechera, Arabidopsis thaliana.

Method synopsis

Creation and analysis of transgenic Arabidopsis lines expressing candidate genes from study species to test gene function and allelic effects. Uses heterologous expression system to isolate gene effects.

Synthesized from method descriptions across 1 paper using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — A gain of function polymorphism controlling complex traits and fitness in nature (2012), Science. Implementations in other papers (listed below) may differ.

1. Create transgenic plants
   Generated transgenic Arabidopsis thaliana lines expressing different BCMA alleles from Boechera stricta, including BCMA1-MT, BCMA2 variants, and BCMA3 constructs under control of CYP79F promoter
   Quantities: Multiple independent transgenic lines per construct, 130 independent transformants analyzedDuration: Plant transformation and generation cyclesConditions: Controlled greenhouse conditions
   Equipment: transformation vectors, plant growth chambers, selection media
2. Measure glucosinolate production
   Analyzed glucosinolate levels in transgenic Arabidopsis plants using established biochemical assays to quantify aliphatic glucosinolates derived from methionine, valine, and isoleucine precursors
   Quantities: Measurements from multiple independent transgenic lines per genotypeDuration: After plant maturation in controlled conditionsConditions: Standardized growth and harvest conditions
   Equipment: HPLC systems, chemical extraction equipment, analytical balances

Typical Equipment

• transformation vectors
• plant growth chambers
• HPLC systems

Output Measurements

• transgenic plant glucosinolate levels
• gene expression patterns
• chemical phenotypes

Papers Using This Protocol (1)

Species Studied With This Protocol