Final Ch. 19

Traditional analysis of mutants (natural or induced) to determine gene function is known as _____.

forward genetics

A powerful genetic engineering technique used for the amplification of sequences of DNA is _____.

polymerase chain reaction (PCR)

Which one of the statements below accurately describes gel electrophoresis?

DNA moves through the gel toward the positive electrode.

TALEN (transcription activator-like effector nuclease)

- In which a protein that normally binds to sequences in promoters is attached to the FOK I restriction enzyme. -The binding protein consists of a series of repeats whose amino acid sequence determines the DNA base sequence that it recognizes.

Vectors (Plasmid Vectors)

-Plasmids, circular DNA molecules that exist naturally in bacteria, are commonly used as vectors for cloning DNA fragments in bacteria. -They contain origins of replication and are therefore able to replicate independently of the bacterial chromosomes.

All of the following are DNA sequencing techniques except _____.

1. DNA fingerprinting 2. Mullis chain-terminating sequencing

These are used as cloning vectors

1. artificial chromosomes 2. plasmids 3. cosmids

Cloning a gene after isolating it requires which of the following?

1. transformation of the recipient cells with the vector with the gene of interest 2. ligation into a vector 3. identification of the transformants that have the gene of interest

CRISPR-Cas System

A molecular tool used for precise editing of DNA that relies on the action of CRISPR RNAs and Cas proteins.

Protein called Cas 9 that cuts DNA, along with two RNA molecules that can be genetically engineered to guide Cas 9 to specific sequences.

CRISPR-Cas 9

What is the key to both ZFN's and TALEN's?

It is the ability to alter the amino acid sequence of the binding proteins in these nucleases in such a way that they bind to a specific target DNA sequence.

How might CRISPR-Cas 9 be used?

It might be used in a similar way to help restore muscle function to boys with Duchenne muscular dystrophy.

Homologous Recombination

May insert the donor DNA sequence into a genome.

Northern Blot

Process by which RNA is transferred from a gel to a solid support such as a nitrocellulose or nylon filter.

RNA in situ hybridization

RNA is visualized while its in the cell. -Method used to determine the chromosomal location of a gene or other specific DNA fragment or the tissue distribution of an mRNA by using a labeled probe that is complementary to the sequence of interest.

R-T PCR

Technique that amplifies sequences corresponding to RNA, reverse transcriptase is used to convert RNA into complementary DNA, which can then be amplified by the usual Polymerase Chain Reaction.

Southern Blot

Technique used to transfer the denatured (single-stranded) fragments from a gel to a solid medium.

Guide RNA

The CRISPR-Cas 9 system can be used to cut and edit the genome. It combines a single guide RNA with a nuclease, which together attach to DNA sequences and make double stranded cuts at specific locations.

Western Blot

The transfer of protein from a gel to a membrane. (Probe= usually an antibody) used to determine the size of a particular protein and the pattern of the protein's expression.

ZFN (Zinc-finger nucleases)

Use a DNA binding domain called a zinc finger attached to a restriction enzyme, most often FOK I. -Most commonly used ZFN's have three of these domains, which together recognize a 9-bp DNA Sequence. -To make double-stranded cuts in DNA, ZFN;s are used in pairs, which increases their specificity to 18 or more base pairs enough to ensure that most genomes are cut only once.

Biotechnology applications include all of the following except _____.

artificial selection for larger fruit size in crops

After separating a complex mixture of DNA fragments according to size, identifying a specific fragment of interest _____.

can be accomplished by Southern blotting

Restriction endonucleases _____.

can be used to create pieces of DNA with cohesive ends

All of the following are required for genome editing with the CRISPR-Cas9 system except _____.

invading DNA protospacers

If researchers have not yet isolated the protein product of a gene, what method can they use to isolate that gene?

positional cloning

Utilizing a genetic library requires all of the following except _____.

positional cloning

In nature, the purpose of the CRISPR-Cas system is to _____.

protect bacteria and archaea from invading DNA elements

Reverse genetics attempts to determine the function of a gene by changing it, using all of the following techniques except _____.

reverse transcription

Recombinant DNA technology is a set of molecular methods used to isolate, manipulate, and study DNA. What does recombinant mean, in this case?

that the DNA used is often derived from two or more sources and combined