Microbiology a clinical approach by tony srelkauskas

Info icon This preview shows pages 23–29. Sign up to view the full content.

View Full Document Right Arrow Icon

Info icon This preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon

Info icon This preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon

Info icon This preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science APPLICATIONS OF GENETIC ENGINEERING Researching gene function and regulation Express recombinant protein in E. coli Regulation of gene expression can be studied by creating a gene fusion Joining the gene being studied and a reporter gene Reporter gene encodes observable trait such as fluorescence GFP, green fluorescent protein Trait makes it possible to determine changes in gene expression ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science DNA SEQUENCING Allows the determination of the exact sequence of nucleotide bases in DNA Knowing DNA sequence of particular cell helps identify genetic alterations Alterations that may result in disease Sickle cell anemia Due to single base-pair change in gene Cystic fibrosis Caused by three-base pair deletion ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science TECHNIQUES USED IN DNA SEQUENCING Dideoxy chain termination method In vitro DNA synthesis reaction requires: Single-stranded DNA template, from which a complementary copy is sythesized Primer that anneals to single-stranded template Dictates the site at which synthesis will initiate DNA polymerase, the enzyme that catalyzes DNA synthesis Each of the four deoxynucleotides (dATP, dGTP, dCTP, dTTP) If these were the only ingredients in the reaction buffer, full-length molecules complementary to the template DNA would by synthesized. Dideoxynucleotides (ddNTPs) Identical to their deoxynucleotide counterparts but lack 3’OH group required for the addition of subsequent nucleotides during DNA synthesis Incorporation causes chain termination Special gel electrophoresis used to separate DNA fragments by size ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science DNA SEQUENCING: PRIMER DNA being sequenced usually has been cloned into a vector. DNA polymerase can only add nucleotides to an existing fragment of DNA, thereby extending the fragment ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science DNA SEQUENCING: CHAIN TERMINATION BY A DIDEOXYNUCLEOTIDE ISBN: 978-0-8153-6514-3 Microbiology: A Clinical Approach, by Tony Srelkauskas © Garland Science Microbiology: A Clinical Approach © Garland Science AUTOMATED DNA SEQUENCING Automated DNA sequencing Most automated systems use fluorescent dyes to detect newly synthesized DNA Gel electrophoresis used to separate fragments into colored bands Laser used to detect color differences Order of color reflects nucleotide sequence of the DNA ISBN: 978-0-8153-6514-3...
View Full Document

{[ snackBarMessage ]}

What students are saying

  • Left Quote Icon

    As a current student on this bumpy collegiate pathway, I stumbled upon Course Hero, where I can find study resources for nearly all my courses, get online help from tutors 24/7, and even share my old projects, papers, and lecture notes with other students.

    Student Picture

    Kiran Temple University Fox School of Business ‘17, Course Hero Intern

  • Left Quote Icon

    I cannot even describe how much Course Hero helped me this summer. It’s truly become something I can always rely on and help me. In the end, I was not only able to survive summer classes, but I was able to thrive thanks to Course Hero.

    Student Picture

    Dana University of Pennsylvania ‘17, Course Hero Intern

  • Left Quote Icon

    The ability to access any university’s resources through Course Hero proved invaluable in my case. I was behind on Tulane coursework and actually used UCLA’s materials to help me move forward and get everything together on time.

    Student Picture

    Jill Tulane University ‘16, Course Hero Intern