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mapping in eukaryotes, mapping in phage, sex determination, chromosome mutations

subunit vaccines consist of one or more surface proteins from the virus or bacteria
Hepatitis B viral vaccine surface protein cloned into yeast expression vector, purified from yeast and packaged for use as a vaccine
Gardasil recombinant capsid proteins provide immunity against HPV
edible vaccines can be produced in food plants. decreas cost of production, avoids needles, no refrigeration required no need for sterile conditions to administer, constant source of recombinant protein
Bt transgenic crops have built in insecticide protection to insect pests
gene testing is one of the first successful applications of recombinant DNA technology
Two examples of prenatal tests amniocentesis and chorionic villus sampling (CVS)
ASO's (allele-specific oligonucleotides) short single stranded fragments of DNA; used as probes to identify different alleles; can differ by as little as one nucleotide (SNPs)
preimplantation genetic diagnosis (PGD) genetic analysis of single cells from embryos created by in vitro fertilization
genotyping microarrays are used to detect mutations in specific genes such as p53 which is commonly associated with cancers
genome scanning allows analysis of DNA for dozens or hundreds of disease alleles
array comparative genomic hybridization (CGH) a microarray technique, identifies copy number variations
genome wide association studies(GWAS) identify genome variations that contribute to disease
gene therapy transferring specific genes into mamalian cells to treat genetic disorders
severe combined immunodeficiency disease (SCID) was the first disease to be treated with gene therapy
structural genomics focuses on the sequencing of genomes, analyzing nucleotide sequences to identify genes/ gene regulatory elements
two methods to genome sequencing 1)clone-by-clone method 2)whole genome shotgun method (shotgun cloning)
clone-by-clone method of genome sequencing generates contigs that are aligned by restriction mapping; overlapping clones are sequenced. organized sequencing of contigs form a restriction map instead of random sequencing and assembly
shotgun cloning method of genomic sequencing genomic DNA cut into contigs. overlapping sequenced fragments aligned using computer programs to assemble an entire chromosome; fragments aligned based on identical DNA sequences
bioinformatics uses computer based approaches to organize, share and analyze data related to gene structure, gene sequence and expression, and protein structure and function
annotation the process of identifying gene, their regulatory sequences and their functions
open reading frames (ORFs) stretches of nucleotides that when translated to protein by computer analysis generate a series of amino acids prior to a stop codon; suggestive of a protein encoding gene
functional genomics study of gene functions based on possible RNAs or proteins they encode;study of regulatory element function
orthologs homologous genes in different species(arise through speciation)
paralogs homologous genes in same species (arise through duplication)
single nucleotide polymorphisms (SNPs) single base changes in the genome; many associated with disease conditions
copy number variations (CNVs) segments of DNA that are duplicated or deleted
ENCODE (Encyclopedia of DNA Elements) created with the aim to use both experimental approaches and bioinformatics to identify and analyze functional elements that regulate expression of human genes
Comparative Genomics compares the genomes of different organisms in order to answer questions about genetics and other aspects of biology
eukaryotic genomes have several features not found in prokaryotes 1)gene density that varies from chromosome to chromosome 2)introns 3)repetitive sequences
transcriptome analysis studies gene expression qualitatively by identifying which genes are expressed and which are not; and quantitatively by measuring the varying levels of expression of different genes
microarray analysis enables simultaneous analysis of all of a sample's expressed genes
proteomics the identification and characterization and quantitative analysis of all proteins encoded by the genome of a cell, tissue, or organism
typical proteomic analysis cells exposed to different conditions, proteins extracted, separated by 2DGE, spots of interest cut out, digested into peptide fragments and analyzed by mass spectrometry
systems biology incorporates data from genomics, transcriptomics, proteomics,etc. to further elucidate components of interacting pathways
Pharmacogenomics the study of how an individual's genetic makeup detemines the body's response to drugs
restriction enzymes bind to a specific recognition sequence (restriction site); cleaves DNA to produce restriction fragments; most of these recognition sequences are palindromic; often cut in a staggered way, leaving "sticky ends"
plasmid an extrachromosomal double-stranded DNA molecule that replicates independently from bacterial chromosomes
transformation the process by which plasmids are introduced using calcium ions and brief heat shock to pulse DNA into cells; electroporation is an alternative to this method
cDNA libraries are prepared by isolating mRNA from cells, synthesizing the complementary DNA using reverse transcriptase, and cloning the cDNA molecules into a vector
a restriction map of cloned DNA segment establishes the number and order of restriction sites and the distance between restriction sites
southern blotting(DNA) is used to find a particular clone in a library, characterize the size of the fragments from restriction digest, determine whether a clone conatains all or part of a gene
Northern blotting (RNA) is used to determine whether a gene is actively being expressed in a given cell or tissue and to study the patterns of gene expressions in embryonic tissues, cancer and genetic disorders
Sanger sequencing the most common method of DNA sequencing. also called dideoxynucleotide chain termination sequencing. it involves the addition of a small amount of one dideoxynucleotide(ddNTP)which causes DNA synthesis to terminate
Next generation sequencing 1)DNA fragments on beads amplified by PCR 2)DNA on beads subjected to pyrosequencing reaction 3)light emitted by pyrosequencing reactions captured by detector
Stages of tumor development 1)cell with genetic mutation 2)hyperplasia 3)dysplasia 4)in situ cancer 5)invasive cancer
clonal orginated from a common ancestral cell that accumulated specific mutations
defects in cancer cells affect genomic stability, DNA repair and chromatin modification
cancer cells have altered DNA methylation patterns
histone modifications are ______ in cancer cells disrupted
apoptosis programmed cell death involving DNA fragmentation, digestion of cellular components, membrane blebbing, and apoptotic bodies being engulfed by phagocytes
the relative concentrations of the ____ and ___ proteins regulate apoptosis Bcl2; BAX
regulation of levels of Bcl2 and BAX can trigger or prevent apoptosis
cancer cells that overexpress Bcl2 are resistant to chemotherapy/radiation
p53 induces the expression of BAX; encodes transcription factor; can arrest the cell cycle at several phases; mutated in more than 50% of cancer cells
proto-oncogenes normally stimulate cell proliferation. encode transcription factors, signal transduction molecules that stimulate cell division and cell cycle regulators
tumor suppressor genes normally inhibit cell proliferation
oncogene mutant proto-oncogene (GOF mutation;dominant); stimulates UNREGULATED cell proliferation
signal transduction initiates a program of gene expression that propels the cell out of G0 and back into the cell cycle. cancer cells often have defects in signal transduction pathways
ras genes encode signal transduction molecules
how does a proto-oncogene become an oncogene? point mutation(ras); chromosomal translocation(CML); amplification (HER2)
oncogenes have two types cellular (c-oncs); viral (v-oncs)
retroviruses can cause cancer by activating a cellular proto-oncogene; carrying an oncogene into a cell;
acute transforming retroviruses infect and transform normal cells into cancer cells (none identified in humans yet)
aneuploidy an organism gains or loses one or more chromosomes
euploidy complete haploid sets of chromosomes are present
polyploidy more than two sets of chromosomes are present
reciprocal translocation involves exchange of segments between nonhomologous chromosomes
Robertsonian translocation breaks at the extreme ends of the short arms of two nonhomologous acrocentric chromosomes
Klinefelter's syndrome male genitalia; XXY
Tuner syndrome single X chromosome; female genitalia
human with three X chromosomes female differentiation; often perfectly normal; sometimes underdeveloped secondary sex characteristics, sterility and mental retardation may occur
human with sex genotype XYY males that tend to be over 6 feet tall
SRY absent in human embryo female differentiation; wolffian duct degenerates; mullerian duct becomes the uterus
SRY present in human embryo male differentiation; mullerian duct degenerates; wolffian duct develops into vas deferens
pseudoautosomal regions (PARs) on Y chromosome have homology with regions on X chromosome
SRY encodes for testes-determining factor which is a transcription factor that acts as a master switch to trigger the formation of testes
primary sex ratio reflects the proportion of males to females conceived in a population
secondary sex ratio reflects the proportion of each sex that is born
dosage compensation balances the dose of X chromosome gene expression in males and females
Barr bodies the inactive X is highly condensed and can be observed in stained interphase cells
Lyon hypothesis X-inactivation occurs randomly in somatic cells
mechanism of inactivation of X X-inactivation center(Xic) is active on the inactive X. it contains the X-inactive specific transcript (XIST) gene. XIST RNA coats the inactive X chromosome
bilateral gynandromorph organisms with one X chromosome lost during the first mitotice division, making them half male and half female in some species
if two viruses simultaneously infect a bacterial cell, could recombination between them occur? yes
intergenic recombination recombination between two loci;
Seymour Benzer produced a genetic map of the rII locus of phage T4
intragenic recombination in phage simultaneous infection of e.coli by two mutant phages produces recombinant WT phages. the frequency is proportional to the distance between the two mutations
deletion mapping provides a rough localization of each mutation. a point mutation is localized in the area of a deletion if it fails to give rise to any WT recombinants in complementation assays
crossing over the reciprocal exchange of chromatid segments between homologous chromosomes; involves the breaking and rejoining of two chromatids)
chiasma place on homologous pair of chromosomes at which exchange is occurring; site of crossing over
map units (also known as centimorgans) are defined as 1% recombination between two genes on a chromosome
interference (in relation to crossing over) reduces the expected number of multiple crossovers when a crossover event in one region of the chromosome inhibits a second event nearby; if interference is positive, there are less double crossovers than expected; if interference is negative,there r more
somatic cell hybridization involves the fusion of two cells in culture to form a heterokaryon; nuclei then fuse to form a synkaryon from which chromosomes are lost at random
heterokaryon a single hybrid cell with two nuclei
ordered tetrad analysis can be used to map the centromere; determines map distance between a gene and the centromeres
unordered tetrad analysis can be used to determine whether two genes are linked on the same chromosome and if so, map the distance between them
Created by: ESPOLADE

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