Moelcular Medicine I Flashcards Preview

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Flashcards in Moelcular Medicine I Deck (48):
1

what the 2 phases of genetic analysis of a disease

find a correlation between amuattion an da diseae
detect or predicting a disease

2

when is DNA sequencing most useful

where the exact nature of the disase causing muation is known

3

limits to DNA sequencing

genes are large and complex, some of the mautions may not be connected to the disease at all

4

indirect methods advantage

require no knowledge about the exact nature of the muation as long as the genomic locus of the mutant allele is known

5

limits to indirect methods

good polymorphic markers are scarce. and you could not be looking in the right place

6

the complete human genome consists of how many basepairs

3 billion

7

WGS

looking at the whole genome

8

WES

only looks at the exons

9

SNP typing

looking just at SNPs which are the areas that are actually different between people

10

wha tis the narrowest approach to sequcning

just look at the SNPs that have been assocaited with a health risk

11

limiations to seuqncing

-doesn't show on which chromsome the muation is
- does not show if they have a deletion of a duplicatio nof a chromsomal region

12

copy number variation (CNV)

observation that chromsomal regions can be present in non-diploid states

13

most CNVs are what

benign polymorphisms

14

how do you do comparative genome hybridization

take sample of single stranded genomic DNA from pt and control. label with die, mybrize to metaphase chromosomes. CNV becomes evident by uneven labeling of template chromsome

15

FISH

allows ID of a chromosomal lobus on a metaphase chromosome by hybridation with a specific flouorescent probe

16

how do you do PCR

1. denature template DNA
2. annel primers
3. polymerase synthesizes complmentary strand

17

how does PCR detect genetic polymorphisms

when using ASOs, only mutant alleles will be amplied
insertions or deletions become immediately obvious

18

how do you do real time PCR

-the amoun tof amplified DNA is measured after each cycle
-determine how many cycles of amplification are required
-high number of templant molecules, fewer cycles will be required to reach the threshold
-templant amount is quantified by the number of cycles required to reach threshold

19

what is reverse transcriptase PCR used for

retroviral load in blood samples (HIV)
expression of oncogenes, and tumor suppressor genes in cancer cells

20

ASOs

tool to detect single nucletotide polymorphisms and mutations

21

what does ASP require

detailed knowledge about the nucletodie sequence of the gene under studyy

22

because oligonucleotide primers are short they anneal only to gerions of ______

high sequence complmentarity

23

mutant allele in aickle cell

AT exchange in codon 6 of beta globin gene

24

DNA microarrays

thousands of oligonucletodie spots printed on an inch paper. use for genetic screening

25

the differences in individual drug responses result from what

genetic polymorphisms

26

goal of pharmacogeomics

find the polymorphisms taht affect drug responses in order to work out an indvidual drug treatment plan

27

what is malignant hyperthermia

rare, authosomal disoder that is the leading cause of deal during anesthesia

28

what happens when someone with malignant hyperthermia goes under anthestia

calcium gets released from muscle cells leading to life threatening episodes of uncontrolled muscel contraction

29

what causes malignant hyperthermia

mutation in ryanodine-receptor (RYR1) gene coding fo rmuscel cell membrane ion channel

30

individual response to warfarin is determined by what 2 enzymes

-gene for detoxifying P450 protein causing slow metabolism of drug
-mutation in vitamin K epoxide reductase

31

polymorphism in gene for detoxifying P450 protein causes what

higher senstivity to warfarin

32

mutation in vitamin K epoxide reductase complex (VKORC1) leads to waht

higher warfarin toelrance

33

how are fetal cells sampled

amniocentesis
chorionic villus sampling

34

amniocentesis

amniotic fluid is withdrawn at 15/16 weeks gestation

35

chorionic villus sampling

cells taken from chorion at 10-12 weeks gestation

36

what condition might chonrionic villus sampling miss

mosaicism

37

the specific bindign of antibodies to antigens is used to what

-detect presence of antigen in sample
-quantify amount of antigen in sample
-localize antigens in vivo

38

what produces antiboties

b lymphocytes

39

how are polyclonal antibodies obtained

challenged an animal with intravenously injected antigen. , then after several days take blood

40

how do you get monoclonal antiodies

-inject mouse with protein
-get spleen cells
-fuse spleen celsl with myeloma cells grown in culture
-select and grow hybrid cells
-select and grow cells making desired antibody

41

hybridoma cells

immortal and produce antibodies

42

what is ELISA commonly used for

detecting PTH
HbA1c
c-reactive protein

43

what do you use sandwich ELISA for

quantify an antigen

44

What do you use indirect ELISA for

presence of antibodies

45

What specific diseases is indirect ELISA used for

HIV
rheumatoid arthritis

46

western blotting

technique that uses specific antibodies to indicate the molecular weight of a protein

47

what is the advantage of western blotting over ELISA

you get better infromation about the antigen

48

what do you use western blotting for

confirm diagnosis of HIV
diagnose parasite infections