Biochem Block 2: Recombinant Dna 3

Front 1

How does DNA chips/DNA microarray work?

Back 1

small chip is dotted with single stranded DNA pieces--each chip represents a different gene. single stranded DNA from the pt is incubated with the chip and a computer analyzes the hybridization.

Front 2

What can be used to determine the allele of a drug-metabolizing enzyme causing the pt to have an adverse reaction to a drug?

Back 2

DNA chips/DNA microarray

Front 3

____________ can be used to determine gene expression.

Back 3

DNA chip or microarray

Front 4

With DNA chip or microarray to determine gene expression, the presence of a tag and its intensity is a _______________ and ______________ analysis of gene expression. This is called ______________.

Back 4

-qualitative
-quantitative
-serial analysis of gene expression (SAGE)

Front 5

To avoid the risk of contamination and minimize sampling errors with SAGE, _____________ is used.

Back 5

laser microdissection of pathological samples

Front 6

SAGE is used in the identification and study of...(5)

Back 6

1. lymphomas (lymphochip array)
2. melanomas
3. breast cancer
4. viral diseases
5. cardiovascular diseases

Front 7

Recombinant DNA techniques can be used to create __________________ using yeast instead of bacteria, eliminating risk of infection.

Back 7

glycosylated antigenic proteins

Front 8

Human growth hormone and human insulin are produced by recombinant DNA using the bacteria ________.

Back 8

e.coli

Front 9

To produce insulin, you need to transform E. coli with one or two plasmids?

Back 9

two: one for the Insulin A chain and one for the Insulin B chain. You will later combine the chains to create active insulin

Front 10

Factor VIII does what?

Back 10

blood clotting. it is transfused to hemophiliac patients. can be produced by DNA recombinant techniques

Front 11

What does TPA do?

Back 11

tissue plasminogen activator is a protease that converts plasminogen to plasmin....plasmin cleaves fibrin to prevent the formation of clots. Treats strokes and heart attacks. can be created with DNA recombinant techniques

Front 12

What does erythropoietin do?

Back 12

hematopoetic growth factor that stimulates production of RBC. can be created with DNA recombinant techniques

Front 13

hematopoetic growth factors produced by recombinant DNA: _______ and _________ are used in bone marrow transplants and after chemotherapy to stimulate WBC production and decrease the risk of infection.

Back 13

Colony stimulating factors (CSFs) and interleukins (ILs)

Front 14

Recombinant ____________ (drug) decreases the frequency and severity of episodes in patients with multiple sclerosis.

Back 14

Beta-interferon

Front 15

What are two transgenic animals used to produce human proteins?

Back 15

goat and sheep

Front 16

Human proteins are produced by the _____________ of the transgenic animal.

Back 16

mammary gland

Front 17

Describe how transgenic animals can be used to make proteins.

Back 17

A gene of interest, which is only active under lactating conditions, is inserted into the nucleus of a fertilized egg. The egg is inserted into the foster mother. Progeny is tested for the transgene. The animals who test positive produce milk that is collected...from which the protein of interest is isolated.

Front 18

Reducing gene expression can be done via what 2 ways?

Back 18

1. degrade mRNA of gene of interest
2. block translation of mRNA of gene of interest

Front 19

Describe the default eukaryotic system for gene silencing.

Back 19

microRNAs (miRNA) is processed as a small double stranded RNA molecule that separates into single strands--these single strands can hybridize with the gene of interest and do one of two things:
1. degrade if pairing is exact
2. inhibit translation when pairing isn't exact

Front 20

If pairing isn't exact, miRNA will ______________.

Back 20

inhibit translation of mRNA

Front 21

If pairing is exact, miRNA will ______________.

Back 21

induce degradation of mRNA

Front 22

Chemically synhtesized miRNA is called...

Back 22

siRNA (small interfering RNA)

Front 23

Genetic screening can be done on...

Back 23

1. prospective parents
2. fetus (with the possibility of medical treatment in utero)

Front 24

When normal genes are introduced into individuals with defective genes, are the defective genes replaced?

Back 24

no, impossible to replace the genes of each and every cell

Front 25

In gene therapy, ________ are used as vectors to introduce genes into human cells.

Back 25

retroviruses

Front 26

What are the limitations of gene therapy?

Back 26

1. only small genes (8 kb) can be used
2. Intercalation of the vector into host DNA may disrupt other genes

Front 27

Human ______________ can also be used as vectors for gene therapy...one advantage of this is that you can introduce large genes (~36 kb).

Back 27

adenoviruses

Front 28

Gene therapy: which vector can only introduce small genes?

Back 28

-retroviruses
-adenoviruses can introduce much larger genes

Front 29

What are the advantages of using human adenoviruses as vectors?

Back 29

-genes are large (36 kb)
-infection does not require division of host cell

Front 30

What are disadvantages of using human adenoviruses as vectors?

Back 30

-Integration of therapeutic genes is not stable in host genome, so expression is only transient and treatment must be repeated periodically
-Complications caused by host immune response to adenovirus

Front 31

In TRANSGENIC ANIMALS, does the introduction of normal genes into somatic genes with defective genes correct defects in the offspring of treated individuals?

Back 31

NO

Front 32

The study/comparison of proteins expressed by normal cells and proteins expressed by a cell under study (tumor cell, cell in a different stage of development) is called...

Back 32

proteomics

Front 33

Proteomics: using _________________ with computer analysis, the upregulation or downregulation of a protein can be determined and specific drug regiments can be formulated as a result.

Back 33

-gel electrophoresis