Devo Lect 3 - techniques Flashcards Preview

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Flashcards in Devo Lect 3 - techniques Deck (20):
1

Gene cloning

Copying genes for study, from cDNA library or genomic

2

Promoter region

RNA poly and TF bind there, control on/off genes, KEY for developmental bio

3

DNA hybridization

complementary binding; southern blots; visualize presence of genes (probes); look for conserved across organisms

4

DNA sequencing

dideoxy nucleotides: fluoresce and terminate

5

northern blot

temporal and spatial (when and where) gene expression patterns; gel, transfer, probe

6

in situ hybridization

temporal and spatial (when and where) gene expression patterns; probe put on tissues, bind to complimentary DNA

7

PCR

amplifies segment of DNA; 95, 37, 72 - denature, anneal, taq polymerase

8

RTPCR

Real time or reverse transcriptase PCR - makes DNA from RNA, then PCR - shows genes that are even barely expressed (sensitive)

9

QPCR

Quantifies the amount of DNA; primer fluoresces so we can measure it by that

10

Expressing a gene to study

in xenopus oocytes, tissue culture, bacteria, transgenic organism and study effects

11

Knock out

insert some DNA to recombine (homologous sequence), get heterozygote, then breed to get homozygous knockout and study effects

12

Antisense DNA

insert little bits of complementary sequences which bind to mRNA, can’t be translated; not used that much

13

RNAi

use a very little bit of dsRNA, stimulates machinery of cell to cut up the RNA of choice

14

Western Blot

where proteins are expressed; need labeled antibody

15

immunofluoresence microscopy

fluorescent antibody so you can see proteins

16

Glofish

Transgenic zebrafish; designed as environmental detector; expresses GFP, RFP, etc. in muscles; insert muscle-specific promoter in front of GFP

17

Bioreactor purposes

A living system to grow large amounts of something, either to isolate something (ie. insulin, vitamins) or a useful protein (ie golden rice)

18

Glofish reproductive rate study

Slower reproduction; due possibly to colour; also due to energy expenditure to express GFP at high levels

19

Aquabounty atlantic salmon

Has chinook salmon growth hormone with promoter from Ocean pout - growth hormone is active all year rather than periodically in wild type

20

Concern about transgenic salmon

Possibly high growth factor in the flesh, and does this have an effect on humans; Process could disrupt normal genes; allergic reactions; competitive advantage over wild type fish?