DNATechnology160-page5 - uptake of DNA into target cells...

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Biotechnology - 5 Recombinant DNA Technology Techniques and Applications In order to make recombinant DNA, a researcher must be able to do the following: 1. The desired gene must be located and isolated (which is why mapping the genome is so important). o Restriction enzymes may used to make fragments of DNA more suitable for analysis. o The DNA probe is one technique used to obtain desired DNA. o Researchers also use complementary DNA (cDNA), which is synthesized from RNA templates, when the DNA sequence can be determined. 2. The target gene or DNA must be made available and in quantities suitable for work. o The DNA polymerase Chain Reaction (PCR) is a technique that amplifies the volume of the desired DNA 3. The target gene or DNA must be transferred to the host and incorporated into the host's DNA and expressed. o Plasmids and viruses are often gene vectors. o Other techniques are also used, including the "DNA gun" and direct
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Unformatted text preview: uptake of DNA into target cells Finding the DNA Source – A Critical Step The desired gene that codes for the protein needed must be located and isolated (which is why mapping the genome is so important). It's not easy, and genes with introns, even if successfully recombined, might not get the mRNA transcripts properly spliced and processed for translation in the host. There are a number of ways in which DNA gets isolated for use. cDNA To solve the intron problem, some genes used for recombinant DNA work are "artificially" made. The DNA made is known as cDNA or copied DNA using a functional mRNA template. Reverse transcriptase enzymes, found in some viruses, can take a strand of mRNA and code a complement DNA strand. The DNA strand can then be used as a template to code its complement to form a double-stranding DNA molecule that can be used to splice into the vector....
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This note was uploaded on 12/29/2011 for the course BIO 151 taught by Professor Edwards during the Spring '10 term at SUNY Stony Brook.

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