HG 10 Genetic Technology and Cloning Flashcards

Flashcards on Cloning, Recombinant DNA Technology, PCR, and Genetic Therapy

Clone

An identical copy of a DNA segment, whole cell, or complete organism derived from a single ancestor.

Cloning

The process of producing identical copies of molecules, cells, or organisms.

Plant Cloning

Cloning plants from single cells or tissues through de-differentiation and re-differentiation.

Artificial Selection

Breeding organisms with desirable traits to get offspring with the best combination of traits.

Embryo Splitting

An artificial 'twinning' process where an egg is fertilized in vitro, the embryo is split into individual cells, and implanted into surrogates.

Nuclear Transfer

The nucleus from an egg cell is removed and replaced with the nucleus from a somatic cell.

Dolly the Sheep

The first cloned animal created using mature adult cells via nuclear transfer.

Gene Cloning

Cloning DNA molecules, not whole organisms, by transferring genes between species.

Recombinant DNA Technology

Technology used to transfer genes between species.

Restriction Enzymes

A way to cut DNA consistently.

Vector

A carrier molecule used to transfer DNA into a cell.

Host Cell

A host cell into which to transfer DNA.

Restriction Enzymes

Bacterial natural defense enzymes that hunt out specific DNA sequences and cut both strands.

Sticky Ends

Recognition sequence is identical on both strands (5’ → 3’).

Ligases

Enzymes that stick overlapping DNA together.

Vectors

Genetically engineered plasmids that carry DNA to the host cell.

Plasmids

Small, circular DNA found naturally in bacteria cytoplasm.

Selectable Markers

Help differentiate between transformed and non-transformed bacteria, usually antibiotic resistance.

Polymerase Chain Reaction (PCR)

Copying DNA sequences for study, targeting only the desired DNA.

Primers in PCR

Short DNA fragments (~20bp) that are complementary binding to DNA flanking the target sequence.

Taq Polymerase

Enzyme for copying that can withstand high temperatures.

Denaturation

Heating dsDNA to 95°C, breaking hydrogen bonds and creating two strands of ssDNA.

Annealing

Primers bind in a complementary fashion to ssDNA at 50-65°C.

Extension

Taq polymerase binds to the 3’ end of the primers at 72°C, reads the DNA base on the single strand, and pulls a complementary nucleotide out of the solution, reforming the double-stranded DNA.

Generate Animal Models for Disease

Genetically engineer animals with disease-causing mutations to study disease processes and transfer findings to human situations.

Create Stem Cells for Medical Treatment

Stem cells have the ability to turn into any cell type or tissue and can be used to repair damaged or diseased tissues.

Bring Back Extinct Species

Requires DNA from the extinct animal and a closely related species to be an egg donor and surrogate mother.

Cloning Livestock

To get better meat, milk, and wool production; a pretty common undertaking.

Plants to Detect Environmental Changes

When the environment changes, the appearance of the plant changes; plants can detect landmines.

Producing Drugs

Generate bacteria that will produce human products, including edible vaccines.

Cloning Pets

Possible with enough money and will, though they don’t always look the same.

Cloning Humans

Raises ethical, legal, and social concerns, though therapeutic cloning is a possibility.

Genetic Therapy

Removing exons with disease-causing mutations, just like the natural process of alternative splicing.

Exon Skipping

A technique that can be used to either remove disease-causing exons from the mRNA or skip over additional exons so that the mRNA sequence makes sense.

Maintaining Reading Frame

mRNA codes for amino acids in 3bp codons

Normal Dystrophin Gene Expression

Provides a structural link to maintain muscle integrity.

Muscular Dystrophy Mutations

Often a result of skipping or deletion of exons.

Exon Skipping Therapy

Artificially skip exons with disease-causing mutations. Skip additional exons to return sequence to the correct reading frame.

Sense and Anti-Sense

Two strands of DNA – Sense (coding) and Anti-sense (template)

Anti-sense Oligonucleotides

Cover the splice recognition site, or bind to exon recognition sites in the exons.

Therapeutic Uses of Cloning

Process of using bacteria to produce human protein products, including edible vaccines, as therapeutics.

Exon Skipping

Technique used to selectively remove exons from an mRNA sequence to bypass disease mutations.

Codon

A sequence of three DNA or RNA nucleotides that corresponds with a specific amino acid or stop signal during protein synthesis.