Step-by-Step Genetic Modification Experiment Illustration

Original Prompt: Followed Protocol (Ito et al., 2012) - isolation method: collecting water samples from the lake and then culturing them to isolate individual bacterial strains. The isolated strains are characterized based on various physical, biochemical and genetic characteristics to determine their identity and characteristics. - Isolation of ddnA Gene: A genomic library of strain KSM1 was constructed and the ddnA gene was isolated from the genomic DNA of strain KSM1 using PCR amplification. - Vector Preparation: The cloning vector pKS13S was employed to host the ddnA gene. - Gene Insertion: The isolated ddnA gene was integrated into the pKS13S vector through molecular cloning techniques. - Verification: Molecular analyses, including restriction enzyme digestion and sequencing, were conducted to confirm the insertion of the ddnA gene into the vector. - Transformation: The recombinant vector, now containing the ddnA gene, was introduced into Escherichia coli cells via transformation, where it underwent replication and propagation. - Binary Vector Construction: The recombinant vector carrying the ddnA gene was transferred into a binary vector compatible with Agrobacterium tumefaciens, facilitating subsequent plant transformation. - Agrobacterium Transformation: Transformation methods such as electroporation were employed to introduce the vector into Agrobacterium cells. - Vacuum Infiltration: The genetically modified Agrobacterium culture, now harboring the ddnA gene, was utilized for vacuum infiltration. Plant material, typically wheat leaves, was immersed in a solution containing Agrobacterium and the ddnA gene construct. - DNA Uptake: The application of vacuum pressure facilitated the penetration of the DNA solution into the plant cells, ensuring efficient delivery of the ddnA gene into the plant genome. - Regeneration and Screening: Following infiltration, the plant tissue was cultured under controlled conditions to promote DNA uptake and regeneration. The infiltrated tissue was systematically screened to identify plants successfully harboring the integrated ddnA gene, achieving the desired genetic modification for enhanced resistance to DON. TIMETABLE GANTT CHART

Model: Imagen 4

Created on 2/17/2024 Report

Updated on 8/6/2025