Biotechnology and Recombinant DNA - PowerPoint PPT Presentation

1 / 18
About This Presentation
Title:

Biotechnology and Recombinant DNA

Description:

4. Put DNA into host cell. Transformation. Electroporation. Protoplast fusion. Microinjection. 5. Now, selection: Fig 9.11. What is being selected? ... – PowerPoint PPT presentation

Number of Views:249
Avg rating:3.0/5.0
Slides: 19
Provided by: wolfw
Category:

less

Transcript and Presenter's Notes

Title: Biotechnology and Recombinant DNA


1
Biotechnology and Recombinant DNA
  • Chapter 9

2
I. Learning Objectives
  • Why genetic engineering?
  • Cloning basics
  • Beyond the basics

3
Overview of DNA manipulation
4
II. Why?
  • A. More efficient, less expensive production
  • B. Genetic information
  • C. Genetic alteration

5
III. Cloning ABCs (Fig 9.1)
6
A. Requirements
  • 1. Target DNA
  • 2. Cloning vector
  • 3. Cut and paste enzymes
  • 4. Put DNA into host
  • 5. Selection procedure

7
1. Target DNA
  • cDNA
  • Genomic DNA

8
2. Vector choice
  • a. Why a vector?
  • b. Plasmids
  • c. Viral vectors
  • d. YACs, MACs and HACs

9
3. Cut and paste enzymes (Fig 9.2)
  • Restriction enzymes
  • Ligase

10
(No Transcript)
11
4. Put DNA into host cell
  • Transformation
  • Electroporation
  • Protoplast fusion
  • Microinjection

12
5. Now, selection Fig 9.11
  • What is being selected?
  • Host grown on selective media

13
III. Beyond the basics
  • A. How do you find your gene of interest?
  • B. A most marvelous work-around cloning
  • C. How to fingerprint a killer

14
A. How do you find your gene of interest?
  • 1. Construct gene library
  • 2. Screen library

15
B. A marvelous work-around
  • 1. Polymerase Chain Reaction
  • Amplification of any target DNA
  • 2. Requires
  • target
  • DNA polymerase
  • primers
  • http//vector.cshl.org/Shockwave/pcranwhole.htm

16
3. Whats it good for?
  • a. Identification of microbe in mixed culture
  • CD-ROM exercise
  • b. Fingerprinting a killer

17
b. Fingerprinting a killer
  • Tracking E. coli 0157H7 (Fig 9.16)

18
Should we or shouldnt we?
  • Safety issues
  • Ethical issues
Write a Comment
User Comments (0)
About PowerShow.com