McGraw Ch.20

steps in cloning a gene

1. Chromosomal DNA is isolated and cut with a restriction enzyme; the plasmid DNA is cut with the same enzyme. 2. The digested chromosomal DNA and plasmid DNA are incubated together. 3. ligation by DNA ligase.

cloning vector

A DNA molecule that acts as a carrier of DNA that is to be cloned is called a(n) ______.

host

A cell that harbors a vector is called a(n) ____ cell

Chromosomal DNA

A particular gene to be cloned is often isolated from

a piece of chromosomal DNA

A recombinant vector contains

Genomic Library

A researcher may use restriction enzymes to digest the DNA of an organism. The fragments of DNA are then ligated individually into many vectors. This collection of recombinant vectors is called a

plasmid

A small DNA molecule that can replicate independently within a host cell and thus make many copies of an inserted gene is called a(n)

copied many times by the host cell

A vector requires an origin of replication so that it can be

recombinant

A vector that contains a piece of chromosomal DNA is referred to as a(n) ___ vector.

Primer extension

During PCR, the process of ___ ___ results when the Taq polymerase catalyzes the synthesis of complementary DNA, starting at the primers.

Restriction Enzymes

Enzymes that bind to a specific DNA sequence and cut the DNA backbone are called

polymerase chain reaction (PCR)

In 1985, Kary Mullis developed a way to copy DNA without vectors or host cells. This technique is called ___ (PCR).

A chain reaction

In PCR, each cycle uses the products of the previous cycle as templates. What do you call this?

Complementary DNA

In PCR, primer extension refers to the synthesis of Blank______ starting at the primers.

Template DNA

In PCR, the DNA to be amplified is called the

DNA

In PCR, the two primers bind to specific sites in the and flank the gene to be amplified.

They are complementary to the flanking sequences.

In PCR, why do the primers bind to specific sites in the DNA on either side of the gene of interest?

short oligonucleotides bind to complementary DNA flanking the gene of interest

Primer annealing occurs when

cut the DNA backbone prior to inserting the DNA to be cloned

Restriction endonucleases are used in gene cloning to

detect and quantify the amount of a specific RNA

Reverse transcriptase PCR can be used to

primers

Short oligonucleotides that flank the region of DNA to be amplified by PCR are called

Origin of replication

The DNA sequence in a vector that allows the replication enzymes of the cell to make lots of copies of the vector

Reverse transcriptase

You have a piece of RNA, and you want to synthesize a complementary strand of DNA. Which enzyme would you use?

PCR reaction steps

denaturation, annealing, extension

The production of new arrangements of DNA

recombinant DNA technology