VLP Key Terms

Virus - like particle

Immunogen: delivery vessel

QBeta

14 kDa monomer

Pentameters'

30nm

180 monomers = polymer

540 lysines

Electroporation

Electrical pulses to create pores —> genetic material permeate membrane

Autoclave

steam sterilization

LB

Lysogeny Broth, nutritionally rich medium

Plasmid

double stranded DNA

code for Kanamycin and VLP

Incubation

uniform temperature and humidity

Streak Plate

progressive dilution of inoculum of bacteria to create single isolated pure colonies

Kanamycin

Aminoglycoside bactericidal antibiotic

Transformation

exogenous genetic material is taken up by a cell through cell membrane

Induction

Bacterial virus transfer DNA along with prophage —> cell breaks apart

IPTG

Cell lysis

breakdown of cell by damage to plasma

chemical and physical means

Lysosyme buffer, DOC, DNAse, Magnesium chloride

Spike

1 colony overnight in LB Kan

OD

Optical Density: light scattered

IPTG

Isopropyl B-D-1 thiogalactopyranoside

allolactose mimic: triggers transcription of the lac operator (RNA operons, access for polymerase type)

Centrifugation

by density

supernatant

soluble liquid reaction: centrifuge, precipitation

Buffer

resist pH change

Lysosyme buffer without DTT

break down plasma —>extraction efficiency

10% deoxycholate

ionic detergent: disrupt, dissociate protein interactions

Sonication

SOUND ENERGY 🔊

Viscosity

resistance to shear forces

want viscous sample = more concentration

2M MgCl2

cofactor: enhance DNA polymerase to boost DNA amp

????

Cofactor

non-protein ions/molecules

help enzymes work

DNAse

purify from prokaryotes

degrades DNA—>hydrolyze into nucleotides (separate)

SCB

gravity column buffer

Sodium Phosphate Dibasic

Gel electrophoresis

separate DNA fragments by size

Agarose gel

check for DNA/RNA presence

SDS gel

Sodium Dodecyl Sulfate

denature protein/break down VLP into monomers

concentration/molecular weight (identify)

Size Exclusion Chromatography

molecular sieve chromatography

size and weight

Gravity Column

purification

glass column+solid phase

Orange loading dye

prepare DNA for agarose

dyes: xylene cyanol FF, orange G

visual tracking

TAE

Tris-acetate-EDTA
Tris base, acetic acid-pH 8.3, EDTA (??)- sequester divalent cations

EDTA: Ethylenediaminetetraacetic acid

agarose to separate DNA/RNA

MES

(2-(N-morpholino)ethanesulfonic acid)

buffer in SDS

highly water soluble

Ammonium Sulfate

precipitate (salt-out) VLPs —>centrifugation

PBS

phosphate buffered saline

pH 7.4

human-body like

dialysis

buffer-exchange

remove unwanted macromolecules

Ethidium Bromide

EtBr

stain DNA in gel electrophoresis

in UV light visible

Ladder

standards of molecule size

Blue Loading Dye

DNA ad samples on SDS

Visual track

Coomassie Blue

stain and identify proteins in SDS

SMPH

bifunctional crosslinker

succinimidyl-6-[9B-maleimidopropionamido)hexanoate]

Apolipoprotein C-1

57 peptides

81 wildtype