Bio225-Chapter 9: Recombinant DNA and Biotechnology

DNA formed by combining genetic material from different sources

What is a recombinant DNA?

The deliverate modification of an organism’s genetic material

What is genetic engineering?

A DNA vehicle used to carry genetic material into a host cell

What is a cloning vector?

Plasmids and viral vectors

What are common cloning vectors?

Cuts DNA at specific recognition sequences

What does a restriction enzyme do?

Short single-stranded overhangs that can base-pair with complementary ends

What are sticky ends?

Seals DNA fragments together by forming phosphodiester bonds

What does DNA ligase do in cloning?

Cut gene and plasmid, join with ligase, transform cells, select or screen, and grow clones

What are the basic steps of plasmid cloning?

Introduction of foreign DNA into a host cell

What is transformation in recombinant DNA technology?

A collection of cloned DNA fragments representing a genome or a set of expressed sequences

What is a DNA library?

A labeled nucleic acid sequence that binds to a complementary target sequence

What is a DNA probe?

A specific protein

What can an antibody probe detect?

Denaturation, primer annealing, and extension

What are the three main steps of PCR?

It survives the high temperatures used during repeated denaturation

Why is heat-stable DNA polymerase used in PCR?

They flank the target region and provide starting points for DNA synthesis

What is the purpose of PCR primers?

Separating molecules based on properties such as size and charge

What is gel electrophoresis used for?

DNA and RNA fragments

What is agarose gel commonly used to separate?

Proteins and small nucleic acid fragments

What is polyacrylamide gel commonly used to separate?

DNA

What does southern blotting detect?

RNA

What does Northern blotting detect?

Proteins

What does Western blotting detect?

Comparing DNA patterns, often through fragment separation and band analysis

What is DNA fingerprinting?

DNA synthesized from an RNA template using reverse transcriptase

What is complementary DNA (cDNA)?

Mature mRNA-derived cDNA lacks introns that bacteria cannot splice out

Why is cDNA useful for expressing eukaryotic genes in bacteria?

An enzyme that synthesizes DNA using an RNA template

What is reverse transcriptase?

Distinguishing likely recombinant plasmids from nonrecombinant plasmids using a color-based indicator

What is blue-white screening used for?

Disruption of the screening gene by an inserted DNA fragment

What does white colony often indicate in blue-white screening?

An intact screening gene and typically a nonrecombinant plasmid

What does a blue colony often indicate in blue-white screening?

Combining cells after their cell walls have been removed

What is protoplast fusion?

A method that delivers DNA-coated particles into cells

What is a gene gun?

Selecting cells that possess a desired vector or genetic trait

What is positive selection in vector cloning?