Isolation of clonable DNA (genomic libraries, cDNA synthesis) cloning vectors (plasmids, bacteriophages, cosmids) plasmid incompatibility and control of copy number. Ligation of DNA fragments, modification of DNA end and different ligation strategies. Direct and indirect methods for the identification of recombinant organisms. Characterization (polymerase chain reaction, nucleic acid sequencing) and mutagenisis of cloned DNA fragments. Gene expression in Gram negative (E.coli) Gram positive (B.subtilis) and yeast cells (S.cerevisea). Use of Agrobacterium and baculoviruses for gene expression in plant and insect cells respectively. Applications in protein engineering, diagnostics and synthesis of useful products.
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