DNA tech. quiz

Produce multiple copies of a single gene and producing abundant protein from that gene.

Discovered in the 1960’s. These enzymes protect bacteria by cutting up foreign DNA at specific sites on a nucleotide sequence.

An enzyme that permanently bonds sticky ends together.

The original plasmid from a bacterium. May contain antibiotic resistance genes. This is the circle of DNA you will add your gene of interest to.

• collection of cloned genes in a bacterium. Ie. Petri dish 1 - colony of bacteria containing the middle of the gene of interest. Petri dish 2 - colony of bacteria containing the end of the gene of interest. Petri dish 3 - colony of bacteria containing the beginning of the gene of interest. In reality genes can be very large and 10s-100s of plates would be needed to house all of the parts of a gene.

Bacterial Artificial Chromosome - a plasmid with many genes. These genes may function together, like the lac or tryp operon genes.

Complementary DNA made from mRNA with reverse transcriptase. No introns in the cDNA. Just exons that actually code for the needed protein.

A plasmid with a strong promoter upstream from the cloned eukaryotic genes. Causes the expression of the gene.

Polymerase Chain Reaction - makes copies of small amounts of DNA.

Process separates DNA by size, by moving it through a gel with electricity. Think of swimming through jello. Smaller pieces move faster than larger pieces.

Gel electrophoresis of DNA, hybridize with a labeled RNA and ssDNA. Allows you to find a gene of interest.

Gel electrophoresis of mRNA, hybridization with labeled RNA and ssDNA. Allows you to find mRNA of interest.

Isolate mRNA, do PCR, run gel. DNA bands on the gel indicate the presence of the mRNA coding for DNA. Ie. B-globin mRNA-a, B-globin mRNA-b, etc.

In the organism

In a test tube or non-living test media.

In the organism

DNA attached to small glass slides or array. Probed with cDNA.

RNA interference - double stranded RNA matches a gene sequence and block translation of mRNA. Knocks down gene expression.

Differentiated cell that can dedifferentiate and form an entire, new adult organism.

Ie. Dolly - sheep that was cloned from a mammary cell nuclei + an enucleated sheep egg. Produced a diploid cell which grew to an embryo.

Single Nucleotide Polymorphism - a single change in every 100-300 nucleotides. Unique to individuals, like fingerprint.

Restriction Fragment Length Polymorphism A SNP in a gene can change a restriction site –> fragments of lengths other than the usual. Ie. CCAATT/CCAATT/CCAATT normal gene sequence –> 3 CCAATT fragments Ie. CCAATT/CCAAGTCCAATT A SNP of T removes one restriction site –> 2 fragments

Short Tandem Repeats - repeated units of 2 - 5 base pairs in specific regions of a genome. The number of repeats varies from person to person and is an identifier like a fingerprint.

Genetically Modified Organism - genes added or removed