Cell Bio Final Terms

Front 1

isoelectric focusing is the

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1. prots can be sep on basis of their rel contents of acidic and basic residues.

-isoelectric point of a prot is the pH at which the net charge is 0.

-separation of prots in a pH gradient gel

-can resolve prots that differ in pI by as little as 0.01 which means that prots tht differ by one net charge can be separated

2.isolectric point focusing can be combined w/SDS PAGE to obtain high resolution separations
-subject a single sample to isoelectric focusing in a horizontal direction
-single lane gel is placed horizontally on top of SDS PAGE slab and are separated in a vertical direction on the basis of mass

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semiconservative

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1. strand separat
2. bind of DNA poly that moves in 5-->3
3. strand syn in 5-->3 direction brings nucleotides into make a newly synthesized strand of template
4. Half of the molecule is conserved and rest is newly synthesized

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isoelectric point of a prot is the

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pH at which the net charge is 0

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plasmid purificat

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enrich plasmid DNA over chromosomal DNA

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purines have

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2 rings

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SDS

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sodium dodecyl sulfate, an ionic detergent that binds to prot giving it a negative charge, unfolds 3º sturcture, solubilizes proteins

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pyrimidines have

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1 ring

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transcription

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DNA->mRNA (nucleus)

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2 D gel electrophoresis

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prots can be sep on basis of their relative contents of acidic and basic residues

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sense

antisense

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mRNA

complementary to the template

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A260/A280 of pure DNA is between

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1.8 and 1.9

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antibodies

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prot molecules produce by beta cells of vertebrates(immunoglobulins) and bind to entities called antigens w/high specificity(light and dark chains)

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A320>5% of A260, there is a presence of

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particulates

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transmittance and concentration are

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inversely proportional

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restriction digests

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loading dye, plasmids, water, restriction enzymes

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nucleotides have

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covalent bonds

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ciliary motion is due to

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bending of ciliary axoneme, as a result of the sliding of the axonemal mts

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quaternary structure

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2 or more indep folded prot chains loosely held together by weak bonds

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plasmid purificat(brief)

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phosphate buffer, SDS, potassium acetate, etOH, Tris/EDTA

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one helical turn in DNA is

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3.4 nm

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bacterial transformat

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induce competence w/ice cold CaCl2, heat shock high temp for bact envel to become susceptible to uptake of DNA, pUC vector placed in competent E.coli cells: mercaptoetOH to competent cells, add pUC plasmid, heat shock, SOC, plate

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G and C have

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3 bonds

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low ratio of A260/A280

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protein

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2º antibody recognizes

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1º antibody coupled w/dye and is where fluorescence is coming from

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DNA extraction

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pronase, NaCl, RNase, etOH, Tris/EDTA

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axoneme

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a bundle of mts(comprise the cilia and flagella), has 9 pairs of outer mts surr by 2 central mts enclosed by a plasma membrane

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tetrahymena are

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fresh water protozoan living in axenic culture

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ionic

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last part has + or - charge

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fluorescent dyes used for

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nuclear morphology

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nucleotides have

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covalent bonds

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electrophoresis

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agarose, buffer, DNA and loading dye, assemble and load samples

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A and T have

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2 bonds

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deciliate the cell by adding

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CaCl2

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polar

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OH groups, NH2 groups

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ethidium bromide

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fluorescent dye that lies betw the DNA bases

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daltons are

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base pairs or kilobases

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A234/A260 show protein contamination when

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>0.5

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nonpolar

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carbon and hydrogens, has SH

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3 kinds of microscopy

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brightfield-wet mount
stereo-cult dish on stage
phase contrast-vary.deg. 1/4 wavelength from Zernike

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DNA is twisted to the

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left and has 2 backbones

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A max

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260 for nucleic acids

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A vs. concentration
(direct linear proportion)

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Beers-Lambert law

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alkaline lysis proced

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cells are pelleted and resuspended in a buffered medium, cells lysed w/SDS, RNase, NaOH, (SDS and NaOH solubilize the prots and elevated pH of NaOH degrades the RNA) alkaline condits denature chromosomal DNA and plasmid DNA separates the strands, salt concentration increases by adding potassium acetate-->chrom DNA, prots, SDS ppt while shorter plasmid renatures, ppted SDS, prots, chrom DNA centrifuged and plasmid in SN is ppted w/EtOH and dissolved in Tris/EDTA

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number of codons

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64

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A320 should be

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<5% A260

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matrix

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polyacrylamide

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histones

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highly basic proteins

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denaturing electrophoresis

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prots denatured in urea, SDS, or a reagent that unfold tertiary struct

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food is engulfed by

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endocytosis-membrane surr. food and sequesters it in a vacuole

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nucleus has a

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fibrous matrix and various DNA binding prots that serve to regulate both transcription and DNA replicat

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cooperative action of highly basic proteins known as histones organize nuclear DNA into

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several chromosomes

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2º structure

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alpha helix and beta plated sheets are held by H bonds and can be a single polypeptide

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A320 shows

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light scattering properties

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native electrophoresis

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separates prots in buffered environmt maintains native charge and configuration

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fluorescent dyes

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rhodamine-red
fluorescein-green

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dimers

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coiled coils locked together and polymerize

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phosphatase

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can prevent annealing of the restriction cut sites and backbone repair by its treatmt

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absorbance

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the logarithm to the base 10 of the reciprocal of the transmittance

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DNA w/concentration of 50 ug/ml has

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A260 of 1

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coiled coil

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amphipathic wraps around another amphipathic

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A min

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234

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spectrophotometer

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an instrument that provides a quantitative measuremt of the absorbed wavelengths

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higher ratio for A260/A280 means

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RNA

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all prots have

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1º and 2º structures

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tetrahymena are

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fresh water protozoan living in axenic culture

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3º structures

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3 D conformat due to folding of polypeptide and interactions of hydrophobic and hydrophilic

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Euk cells have

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complex cytoplasmic membrane systems like ER and membrane compartmts like mitochrondria

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electrophoresis

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principle that charged molecules migrate in an electric field

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cytoskeleton-netwk of interlock. structs consist of

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microtubules, microfilaments, intermediate

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amphipathic

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hydrophobic aa's in helix spaced 3 or 4 posits apart in a sequence

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indirect immunofluorescence

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binding of secondary to primary enables us to see fluorescent image

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1º struct

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a.a in polypeptide chain w/peptide bonds

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cytoskeleton contains many

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basal bodies arranged in longitudinal row, each w/attached cilium, located near a complex called oral apparatus

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translation

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mRNA-tRNA-prots (ribosomes)

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globular prots are

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tertiary structures

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direct immunofluorescence using

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DAPI(4'6 diamido 2 phenylindole)
-used on nonliving cells since it cannot penetrate the plasma membrane of living cells, therefore etOH is needed to fix the cells and then label them w/DAPI

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transmitted light generates a

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electric current when it strikes a phototube

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1 or more nucleoli define

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intense regions of RNA synthesis

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photoelectric tube

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is connected to a galvanometer and equates current w/% transmittance