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Flashcards in molecular bio lab Deck (15):
1

gene

a length of DNA that codes for a biologically useful product

2

plasmids

extra chromosomal genetic elements
- double stranded circular DNA molecules
-replicate independent of chromosomal (allows for amplification)
- carry non-essential genes
- can be natural or engineered, can carry antibiotic resistant genes

3

pGlO

carries three genes and an ori (origin of replication)

4

GFP

green fluorescent protein, the gene we are cloning

5

araC

a regulatory protein, links GFP expression to presence of arabinose

6

bla/ beta-lacatamase

an enzyme that breaks down ampicilin and makes plasmid-bearing cells antibiotic resistance

7

ori

origin of replication
- a region of plasmid DNA where proteins bind to initiate DNA replication (needed to make more copies of the plasmid in cells)

8

PCR steps

1. denature template strands by heating, strand separate (95 degrees)
2. Cool to let primers anneal; primers bind (55 degrees)
3. allow polymerase to extend to primers; nucleotides add to 3' end of primers

repeat cycle 25-30 times resulting in a billion fold increase in source DNA

9

genetic engineering

-the ability to move and join pieces of DNA
- purposes: study of gene, purification of protein, alteration of plants, animals
- need enzymes to cut and paste DNA, and a carrier (vector) for DNA

10

Restriction Endonucleases

- bacterial proteins which bind DNA at a specific site and cleave a phosphodiester bond
- recognition site: usually 6 bp and palindromic, protein binds a dimer

11

DNA ligase

- enzyme that makes phosphodiester bonds between pieces of DNA
- normally joins okazaki fragments during DNA replication

12

vectors

- a gene by itself will not replicate on its own within a cell
- must be attached to something with replication controls= could be chromosomal ori or disabled virus

13

cloning

-cutting and pasting allows for recombinant DNA
- cloning is making large amounts of identical copies

14

getting DNA into a cell to clone

transformation- uptake of naked DNA
transjection- use of a viral vector
injection- using a fine glass needle
electroporation- electric shock makes pores
projectiles- gold and tungsten beads covered in DNA

15

agarose gel electrophoresis

- DNA is electronegatively charged so it migrates to positive charge
- smaller fragments of the DNA go through faster and farther