11 Questions
What should be preserved in the coding sequence or ORF?
Start codon and stop codons
What is the function of a promoter sequence?
It is located upstream of the coding region and is essential for transcriptional activation.
What are the two types of promoters that may be present?
Constitutive or inducible promoter
How can gene expression be improved by the addition of intronic sequences?
By addition of intronic sequences like the Catalase gene intron.
When is it necessary to alter codons in a gene sequence?
It is sometimes necessary when optimizing codon usage for different organisms.
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 Event Selection.
What are the essential requirements in a gene transfer system for the production of transgenic plants?
a) the target DNA, b) availability of a target tissue, c) a method to introduce DNA, d) a procedure to select and regenerate transformed plants.
What does the isolated DNA sequences used in plant transformation contain?
Isolated DNA sequences contain the gene of interest and other non-genic regions, some of which may have regulatory roles.
What is another term for plant transformation?
Plant genetic engineering or bioengineering.
What are the different methods available for gene isolation in plant transformation?
Methods like isolation from DNA libraries, direct amplification by RNA/DNA PCR.
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.
- It is the predictable alteration of the plant genome by specific incorporation of DNA sequences using recombinant DNA technology.
- Also known as plant genetic engineering or bioengineering.
Three-Step Process of Plant Transformation
- Step 1: DNA Delivery to Target Cells
- Step 2: Selection and Regeneration Event
- Step 3: Expression of the Introduced Gene
Requirements for a Plant Transformation System
- Target DNA
- Availability of target tissue including cells competent for plant regeneration
- A method to introduce DNA into those regenerable cells
- A procedure to select and regenerate transformed plants at a satisfactory frequency
DNA Gene Isolation
- Methods: isolation from DNA libraries, direct amplification by RNA/DNA PCR
- Isolated DNA sequences contain gene of interest and other non-genic regions
- Unwanted sequences are trimmed from the N' and C' terminals, 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
- Promoter: a sequence upstream of the coding region (5') that can be constitutive or inducible
- Terminator: can be artificially added (e.g. NOS) or existing terminator sequences/polyA tails preserved
- Introns: addition of intronic sequences may improve gene expression (e.g. catalase gene intron improves gus expression)
- Codon optimization: alteration of codons necessary for optimal gene expression in certain organisms (e.g. prokaryotic genes, animal genes)
- Boundary sequences: attaching certain sequences improves stability of the gene expression
Explore the process of plant transformation, which involves introducing exogenous genes into plant cells using recombinant DNA technology. Learn about the predictable alteration of plant genomes through the specific incorporation of DNA sequences.
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