· Introduction to plasmids - structure, high
and low copy
number, different forms, plasmid encoded phenotypes.
· Replication of plasmids - direction of replication,
priming, elongation, termination, control of replication,
host functions, incompatibility, amplification.
· Recombination and transposition - homologous
recombination, RecA, crossing over, IS elements,
transposons, inverted repeats, target duplication, frequency
of transposition, model for transposition, rearrangements,
transposon mutagenesis.
· Bacteriophages - phage l, lytic and lysogenic life cycle,
replication of l, packaging, control of lysogeny, phage M13
life cycle and replication.
· Recombinant DNA technology - restriction enzymes, joining
DNA molecules, plasmid, phage, cosmid and phagemid vectors
for gene cloning, introduction of DNA into cells, gene
libraries, identification of recombinant clones, the
polymerase chain reaction, DNA sequencing, DNA synthesis,
site directed mutageneis, RNA analysis.
· DNA sequencing methodologies.
· In vitro site-specific mutagenesis.
· Methodologies for studying protein-nucleic acid
interactions.
· Procaryotic biosynthetic operon (tryptophan operon) -
molecular mechanism of control.
· Eucaryotic genomes (with emphasis on the human genome),
structural features, repeated vs unique sequences,
organisation and distribution of genes/gene families, gene
structure, pseudogenes.
· Chromatin structure, DNA methylation in eucaryotes,
Nucleosonal phasing.
· Overview of gene expression. RNA polymerises, RNA
transcription/processing (including RNA splicing
mechanisms). Differential/tissue specific expression,
enhancers. Translation, mRNA stability, post-tranlational
processing and protein structure.
· Organisation and expression of immunoglobulin genes, DNA
splicing, class switching, allelic exclusion, clonal
selection, somatic mutation.
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Essential:
7 Molecular Genetics of Bacteria', J. Dale (Wiley).
7 'Molecular Biology', D. Freifelder (Jones & Bartlett).
7 'Principles of Gene Manipulation', Old & Primrose, 5th Edition (Blackwell).
7 'Genetics - a Molecular Approach', T.A. Brown, 2nd Edition (Chapman & Hall).
7 'Recombinant DNA', Watson, Gilman, Witkowski and Zoller (Scientific American Books, 1992).
Supplementary:
7 'Bacterial Plasmids', K. Hardy (van Nostrand).
7 'Human Molecular Genetics', Strachan and Read (BIOS Scientific, 2000).
7 'An introduction to GENETIC ANALYSIS', Griffiths et al.
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