11 Questions
What is the purpose of trimming unwanted sequences in the N’ and C’ terminals?
Preserve the coding sequence or open reading frame (ORF)
What does the coding sequence contain in terms of start and stop codons?
Start codon (ATG) and stop codons (TAA, TGA, TAG)
What is the role of a promoter in transcriptional activation?
A promoter sequence should be at the upstream of the coding region (5’). It may be constitutive or inducible.
Why may introns be added to a gene sequence?
To improve gene expression
Why is codon optimization important in gene editing?
AT/GC content varies among organisms, leading to preferences in codon usage.
What is plant transformation?
Plant transformation is the predictable alteration of the plant genome by specific incorporation of DNA sequences.
What are the three steps involved in the introduction and expression of genes into plants?
DNA Delivery to Target Cells, Selection, and Regeneration.
What are the essential requirements in a gene transfer system for production of transgenic plants?
a) the target DNA, b) availability of a target tissue including cells competent for plant regeneration, c) a method to introduce DNA into those regenerable cells, d) a procedure to select and regenerate transformed plants at a satisfactory frequency.
How is gene isolation achieved in plant transformation?
Gene isolation can be done through methods like Isolation from DNA libraries or direct amplification by RNA/DNA PCR.
What is another term used to refer to plant transformation?
Plant genetic engineering or bioengineering.
What is the main goal of plant transformation?
The main goal is to introduce and express specific genes in plants.
Study Notes
Plant Transformation
- Plant transformation involves introducing exogenous genes into plant cells, tissues, or organs using direct or indirect means developed by molecular and cellular biology.
Definition
- Plant transformation is the predictable alteration of the plant genome by specific incorporation of DNA sequences using recombinant DNA technology.
- It is also known as plant genetic engineering or bioengineering.
Process of Plant Transformation
- The introduction and expression of genes into plants is a three-step process:
- DNA delivery to target cells
- Selection and regeneration event
- Selection requirements/components of a plant transformation system
Essential Requirements
- Target DNA
- Availability of target tissue, including cells competent for plant regeneration
- Method to introduce DNA into those regenerable cells
- Procedure to select and regenerate transformed plants at a satisfactory frequency
Gene Isolation
- Different methods are available, such as isolation from DNA libraries and direct amplification by RNA/DNA PCR
- Isolated DNA sequences contain the gene of interest and other non-genic regions
- Unwanted sequences are trimmed, preserving the coding sequence or open reading frame (ORF)
Coding Sequence
- Contains a start codon (first ATG) and stop codons (TAA, TGA, TAG)
Transcriptional Activation
- Requires promoter, terminator, and intron sequences
- Promoter: a promoter sequence is required upstream of the coding region (5’)
- Terminator: terminator sequences may be artificially added (e.g. NOS) or existing terminator sequences/PolyA tails may be preserved
- Introns: addition of intronic sequences may improve gene expression (e.g. Catalase gene intron improves GUS expression)
Codon Optimization
- AT/GC content of DNA varies with the organism and may require alteration of codons (e.g. prokaryotic genes, animal genes)
- Boundary sequences: attaching certain boundary sequences sometimes improves the stability of the gene
Test your knowledge on plant transformation, genetic engineering, and the alteration of plant genomes through the incorporation of DNA sequences. Learn about the techniques and methods used in plant biotechnology.
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