Recombinant DNA Technology

By the late 1970s, techniques for easily sequencing ______, synthesizing oligonucleotides, and expressing eukaryotic genes in prokaryotes had also been developed.

Between 1983 and 1985, Kary Mullis developed a new technique to synthesize large quantities of a DNA fragment without ______ it.

This is the polymerase chain reaction or ______ technique.

The first step in the PCR technique is to ______ the DNA at 94°C for 15 seconds.

The third step in the PCR technique is ______ at 68°C for 60 seconds.

The easiest vectors to work with are ______.

RDNA phages and other viruses are more conveniently stored for long ______.

Larger pieces of DNA can be cloned with cosmids and artificial ______.

______ is synthesized for the treatment of diabetes.

Erythropoeitin is used in the treatment of ______.

Diagnosis of ______ disease can be done using genetic engineering.

Howard Temin and David Baltimore discovered the enzyme called ______.

The restriction enzyme ______, isolated from E. coli, cleaves the DNA between G and A in the base sequence GAATTC.

The EcoRI enzyme cleaves DNA leaving two ______ ends.

Reverse transcriptase can be used to construct a DNA copy called ______.

The restriction enzyme EcoRI cleaves the DNA between G and ______ in the base sequence GAATTC.

The mRNA strand ends with a ______ tail.

Once fragments have been isolated, they are ligated with an appropriate ______, such as a plasmid.

The first step is to cut the plasmid with the restriction enzyme ______.

The plasmid and donor DNA are joined together by the enzyme ______, forming a recombinant DNA molecule.

The ______ DNA is also cut with the restriction enzyme EcoRI, creating complementary sticky ends.

The recombinant DNA molecule is introduced into a ______ cell, which is then grown in a medium containing tetracycline.

One of the easiest approaches is to cut the plasmid and donor DNA with the same ______ enzyme.

Cells that have integrated the recombinant DNA molecule will be resistant to ______ and will form colonies on the petri dish.

This molecule can be transformed into a bacterial cell, where it can be transcribed and ______.

The next milestone was the successful generation of recombinant DNA ______.

They allowed the sticky ends of fragments to anneal then covalently joined the fragments with the enzyme DNA ______.

Within a year, plasmid ______, or carriers of foreign DNA fragments during gene cloning, had been developed and combined with foreign DNA.

DNA cut by a restriction ______.

Discovery of Southern blotting technique for detecting specific DNA fragments so that a particular gene could be ______ from a complex DNA mixture.

The Southern blotting technique depends on the specificity of base ______ in nucleic acids.

They normally protect the host cell by destroying phage DNA after its ______.

Cells protect their own DNA from restriction enzymes by ______ nucleotides in the sites that these enzymes recognize.

Incoming foreign DNA is not methylated at the same sites and often is ______ by host restriction enzymes.

There are ______ general types of restriction enzymes.

Types I and III cleave DNA away from ______ sites.

Type II restriction endonucleases cleave DNA at specific ______ sites.

The type II enzymes can be used to prepare DNA fragments containing specific ______ or portions of genes.

EcoRI is a restriction enzyme derived from ______ coli that recognizes the sequence 5’-G-A-A-T-T-C-3’.

One can extract all the DNA from an organism, cleave the DNA into ______, isolate the fragment of interest for cloning.

All of the fragments can be cloned by means of a suitable ______, and each clone tested for the desired gene.

A DNA fragment can directly be ______.

Agarose or polyacrylamide gels are used to separate DNA ______.

The deliberate modification of an organism's genetic information by directly changing its nucleic acid genome is called ______.

DNA molecules are placed in an electrical ______ and allowed to migrate toward the positive and negative poles.

By inserting recombinant DNA molecules into an organism, these molecules can be ______.

The molecules separate because they move at different rates due to their differences in ______ and size.

Recombinant DNA technology opens up totally new areas of ______ and applied biology.

Negatively charged DNA fragments move toward the positive electrode or ______.

One of the first breakthroughs leading to recombinant DNA technology was the discovery of microbial enzymes called ______.

Each fragment’s migration rate is inversely proportional to the log of its ______ weight.

These enzymes recognize and cleave specific sequences about ______ base pairs long.

Recombinant DNA is DNA with a new sequence formed by joining fragments from two or more different ______.