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RECOMBINANT DNA AND GENOMESRECOMBINANT DNA AND GENOMES Molecular cloning (vs. cellular like Dolly) copies of 1 gene at a time Genetic engineering: making/using recombinant DNA discovery of 2 very important things: small circular DNA molecules called plasmids that replicate in bacterial cells especially prevalent in resistant microbes enzymes that cut double-stranded DNA at specific nucleotide sequences = restriction enzymes along with DNA ligase, these 2 things make it possible to recombine DNA molecules in the lab 2 sets of DNA molecules combined = recombinant DNA molecule restriction enzymes make “staggered” cuts in the vector DNA “sticky ends” – small regions of single strands that can base-pair with their complimentary sequences restriction sites are often palindromes DNA ligase used to make covalent bonds between insert DNA and vector DNA transformation: insertion of recombinant DNA into cells selection: only cells that have received a plasmid can grow in the presence of the antibiotic plasmid contains a selectable marker libraries = collections of recombinant DNA genomic libraries contain clones representing all of the DNA sequences in a genome each bacterial cell used to clone contains a different recombinant DNA molecule in some cases, you would not want to clone genomic DNA (contains introns – only want the 1.5% expressed) you would make cDNA, starting with mRNA from a specific organ or tissue reverse transcriptase (special DNA pol) from retroviruses to copy mRNA to DNA once cDNA is double-stranded by a regular DNA pol it can be cloned into vectors clones represent genes that are expressed recombinant DNA molecules can be introduced into plants and animals GMO or transgenic organism: contains recombinant DNA concerns about GMOs consumption/exposure could cause allergic reactions or toxicity insect resistance could kill both helpful and destructive species are we making transgenic humans? gene therapy is a transient kind of genetic modification recombinant DNA does not become part of the genomeGENOMESHuman genome: 1.5% exons (giving rise to proteins, rRNA or tRNA) ~300 bases (introns ~3000


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CSU BC 351 - RECOMBINANT DNA AND GENOMES

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