r-Dna technology

Bacterial Transformation (Chemical/Electroporation)

Introduction of plasmid DNA into bacterial cells using either chemical competence (CaCl₂) or electric pulses (electroporation).
 

Phage-Mediated Transduction

Transfer of genetic material between bacteria using bacteriophages as vectors.
 

Gene Cloning & Plasmid Vector Construction

Insertion of target genes into plasmid vectors for amplification and expression.
 

Recombinant Protein Expression in E. coli / Yeast

Production of heterologous proteins in microbial hosts for research or industrial use.
 

Purification Using Affinity Chromatography (e.g., Ni-NTA)

Isolation of recombinant proteins tagged with His, GST, or other affinity markers using specific resins.
 

Site-Directed Mutagenesis

Introduction of precise mutations into a DNA sequence to study protein function or gene regulation.
 

CRISPR/Cas9 Genome Editing

Targeted modification of genomes using CRISPR-Cas9 technology for knockout, knock-in, or gene correction.
 

Reporter Gene Assays (GFP, LacZ)

Use of reporter genes to monitor gene expression, promoter activity, or cellular events.
 

Gene Knockout or Knock-In Models

• Knockout: Deleting a gene to study loss-of-function.

• Knock-in: Inserting/modifying a gene to study gain-of-function.
   

Construction of Expression Vectors

Designing plasmids with promoters, selectable markers, and tags for efficient gene expression.
 

Screening of Recombinant Colonies

Identifying bacterial colonies carrying the desired recombinant plasmid using blue-white screening or PCR.
 

Use of Selectable Markers (e.g., Antibiotic Resistance)

Ensures survival of only those cells containing the recombinant plasmid/vector.
 

Use of Shuttle Vectors in Multiple Hosts

Plasmids engineered to replicate in more than one host species (e.g., bacteria and yeast).
 

Fusion Protein Design (e.g., His-Tag, GST-Tag)

Engineering proteins with tags to simplify purification and detection.
 

Epitope Tagging and Detection

Adding small peptide sequences to proteins to facilitate detection by antibodies.
 

Cloning of Synthetic Genes

Insertion of artificially designed or synthesized DNA sequences into expression systems.
 

Development of Biosensors Using Recombinant Technology

Design of genetically engineered microbes or proteins for detection of toxins, pollutants, or biomolecules.
 

RNA Interference (RNAi) Constructs

Design of shRNA or siRNA constructs to silence specific genes at the mRNA level.
 

Lentiviral/AAV-Mediated Gene Delivery

Use of viral vectors to deliver genes into mammalian cells for stable expression or therapeutic purposes.
 

Recombinant Vaccine Development

Engineering recombinant proteins or viral vectors for safe and effective vaccines.