nestors microbio chapter 9

Biotechnology

The use of microbiological and biochemical techniques to solve practical problems.

Recombinant DNA Techniques

Methods used to genetically alter organisms by isolating, modifying, and transferring genes.

Genetic Engineering

The process of altering an organism's genetic material to produce valuable proteins, specific DNA sequences, or study gene function.

DNA Sequencing

Determining the exact order of nucleotides in DNA to decipher amino acid sequences and study genetic relatedness between organisms.

Polymerase Chain Reaction (PCR)

A technique used to amplify a specific segment of DNA millions of times in a few hours for diagnosis or forensic analysis.

Probe Technologies

Techniques such as colony blots, FISH, and DNA microarrays used to detect specific DNA sequences or monitor gene expression.

Restriction Enzymes

Enzymes isolated from bacteria that recognize specific nucleotide sequences and cut DNA at defined sites.

Palindrome (in DNA Recognition)

A sequence that reads the same in the 5' to 3' direction on both complementary strands of DNA.

Restriction Fragments

Short pieces of DNA produced when restriction enzymes cut a larger DNA molecule.

Cohesive Ends (Sticky Ends)

Staggered single-stranded ends of DNA created by restriction enzymes that can anneal to complementary ends.

DNA Ligase

An enzyme that forms covalent bonds between adjacent nucleotides, permanently joining DNA fragments together.

Restriction Enzyme Naming Convention

Named after the genus, species, and strain of the bacterium from which the enzyme was originally isolated (e.g., EcoRI from Escherichia coli RY13).

DNA Cloning

Isolating a specific DNA fragment, inserting it into a vector, and introducing it into a host cell to produce identical copies.

Cloning Vector

A modified plasmid or bacteriophage used to carry foreign DNA into a host cell where it can replicate independently.

High-Copy-Number Vector

A vector plasmid present in large numbers within a host cell, leading to high yields of gene copies and protein production.

Applications of Recombinant DNA Technology

Producing pharmaceutical proteins (insulin), vaccines, industrial enzymes (chymosin for cheese), and herbicide-resistant crops.

Reporter Gene

A gene attached to a regulatory sequence of interest that encodes an easily observable product to monitor gene expression.

Green Fluorescent Protein (GFP)

A popular reporter protein that fluoresces green under specific light when the fused target gene is expressed.

Transgenic Organisms

Eukaryotic plants or animals that have been genetically engineered to contain foreign genes.

Ti Plasmid

A plasmid from Agrobacterium tumefaciens used as a vector to introduce foreign genes into plant cells.

Glyphosate (Roundup) Resistance

A genetic engineering modification that allows crops (soybean, cotton, corn) to survive treatment with biodegradable herbicide.

Origin of Replication (ori)

A sequence in a plasmid vector required for the plasmid to replicate independently within the host organism.

Selectable Marker

A gene in a vector (such as ampicillin resistance) that allows only cells containing the vector to grow on selective media.

Multiple-Cloning Site (MCS)

A region in a vector containing recognition sequences for several different restriction enzymes.

Second Genetic Marker

A gene in a vector (such as lacZ') used to distinguish cells containing a recombinant plasmid from those with an intact (empty) plasmid.

Genomics

The field of biology focused on sequencing, analyzing, and comparing full genomes of organisms.

Bioinformatics

The interdisciplinary field that develops computational tools to store, organize, and analyze massive amounts of biological sequence data.

Human Microbiome Project

An initiative using DNA sequencing technologies to determine the biological diversity of normal human microbial communities.

Essential Components for PCR

Target DNA template, DNA polymerase (e.g., Taq polymerase), primers, and deoxynucleotides (dNTPs).

DNA Microarrays

Solid supports arrayed with thousands of short DNA probes, used to hybridize labeled cDNA and study global gene expression levels.