Authentic knowledge.

1. Transgenic Crops and their Molecular
Analysis
M.PHOOL BADSHAH

2. Introduction
• Transgenic crops are those which are genetically inherited by
foreign gene.
• Transgene which are inserted into plants for better traits and
varieties.
• Several methods are involved for inserting foreign gene and
their molecular analysis.
• First of all gene isolation, gene transfer into host plant by
desired vector such as agrobacterium gene transfer system.
• Transformed plant are cultivated into sterile environment
condition with proper nutrients and hormones.

3. Genetically Modified Crops
• Isolation gene of interest and their
modification.
• Genetic transformed via desired vector.
• Tissue culture and regeneration (organogenesis
& embryogenesis).
• Selection & Screening.

4. Molecular Analysis of Transgenic Crops
• Several methods are involved:
i. PCR:
 It amplifies the specific region of plant DNA, through PCR
we can detect foreign gene or gene of interest in plant.
Confirmation occur through PCR amplification. If not
amplified then no foreign gene are present in plant or inhibit
their expression.

5. ii. Southern Blotting:
 It is used to analyze the integration pattern of transgene in
plant species.
 It contain nitrocellulose membrane which is firstly contain
DNA which is digested and bands appear on gel
electrophoresis.
 It contains probes for proper detection.
 Southern blotting finally tell us the transgene integrity.

6. iii. Northern Blotting & RT-PCR:
 In this technique RNA transcript are used for seeing
expression.
 RT PCR amplify and detects the RNA transcript and finally
showing the gene expression.
 Northern blotting involves the separation of RNA molecules
on gel and use membrane with specific probe for detection.
 These techniques are usually detect the Gene expression.

7. iv. Western Blotting:
 It is usually involved for expression or presence of protein
incoded by transgene.
 It usually involved the protein extraction, separation on gel,
transfer to membrane, and detection using specific
antibodies.
 Nitrocellulose membrane are used for detection of protein
and specific antibodies.

8. • These molecular analyses provide valuable
information about the presence, integration, stability,
and expression of the transgene in the genetically
modified plants.
• Helping to evaluate their suitability for desired traits
and subsequent applications.