Human Molecular Genetics - Recombinant DNA Technology

Cloning Vectors
- Hold DNA fragments, increase copies via replication. Types include:
- Plasmids, Bacteriophages, Yeast Artificial Chromosomes (YAC).
Expression Vectors
- Contain inducible promoters for controlled gene expression to avoid disruptiveness in host cells.
Inducible Promoters
- Active only when a specific inducer is present (e.g., lac promoter in E. coli).
Restriction Enzymes
- Molecular tools that cut DNA at specific sites, essential for recombinant DNA construction.
- Recognition sequence characteristics have probabilistic occurrences in genomes.
Cloning Process
- Involves cutting DNA, inserting fragments into vectors, and sealing with ligases.

Genomic Libraries
- Represent entire genomes via random DNA fragment cloning. - Include exons, introns, and regulatory elements.
cDNA Libraries
- Produce complementary DNA using mRNA as a template.
Screening Techniques
- Utilize hybridization and specific probes to identify genes of interest in libraries.

Southern Blotting
- Used for DNA analysis to confirm gene integration in modified organisms. - Steps include: DNA digestion, gel electrophoresis, and hybridization with probes.
Yeast Two-Hybrid
- Assesses protein-protein interactions using genetically modified yeast to assess transcription factor functionality. - Common reporter genes: HIS3, LacZ.

Single Nucleotide Polymorphisms (SNPs)
- The primary source of human genetic diversity, occurring about once every 100-300 bases in the genome.
- Methods of identification include PCR amplification and DNA sequencing techniques such as the Sanger method.

Understanding the requirements and processes for DNA manipulation via vectors, restriction sites, and gene expression.
Familiarization with library screening, SNP identification, and techniques for studying gene interactions.