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Biochemistry > RNA: Molecular Genetics > Flashcards

RNA: Molecular Genetics Flashcards

0
Q

DNA containing sequence derived from two different species.

A

Chimeric DNA

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1
Q

Altered DNA that results from the insertion of a sequence of depxyribonucleotides not previously present into an existing molecule of DNA.

A

Recombinant DNA

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2
Q

A labelled oligonucleotide that is used to identify a specific base sequence in a target DNA.

A

Probe

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3
Q

Association of complementary strands of nucleic acids.

A

Hybridization

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4
Q

Based on movement of charged particles in an electrical field toward an electrode of opposite charge. Oliginucleotide fragments are negatively charged at neutral pH because of the presence of _______ groups. Separation based in size with smaller fragments moving faster than the larger fragments.

A

Gel Electrophoresis. PO4 -2.

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5
Q

Types of Gel E.: for separation of larger fragments (thousand of oligonucleotides)

A

Agarose

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6
Q

Types of Gel E.: for smaller fragments (hundreds of oligonucleotide)

A

Polyacrylamide

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7
Q

Radioactive labeling with 32por 35s. After separation, labeled oligonucleotides placed in contact with an X-ray film; will show labeled substances as dark bands. Labeling with fluorescent compound like ___________.

A

Autoradiography. Ethidium bromide.

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8
Q

Used to identify DNA with specific base sequences . Takes advantage of property of nitrocellulose that is tenaciously binds single-stranded DNA.

A

Southern Blotting

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9
Q

A labeled fragment of nucleic acid containing nucleotide sequence complementary to a gene or genomic sequence that one wishes to detect in a hybridization experiment.

A

Probe

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10
Q

Detection of specific RNA sequences

A

Northern Blotting

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11
Q

Detection of specific proteins through use of antibodies

A

Western Blotting

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12
Q

Enzymes that cut DNA at highly specific sequences within the molecule. Protect the host bacterial DNA from DNA from foreign organisms. Always exist with site-specific DNA ________ that methylates the host DNA rendering it unsuitable substrate for digestion by restriction enzymes.

A

Restriction Endonucleases. Methylase.

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13
Q

Restriction Endonucleases is named after the bacterium from which they were isolated

A

EcoRI

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14
Q

Types of Restriction E.: cleaves DNA about 25 bp from the recognition sequence.

A

Type III

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15
Q

Types of Restriction E.: cleaves DNA at random sites that can be more than 1,000 base pairs from the recognition sequence.

A

Type I

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16
Q

Types of Restriction E.: cleaves the DNA within the recognition sequence itself; requires no ATP recognition sequences are usually 4-6 bp long and palindromic.

A

Type II

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17
Q

Types I & III are generally large multisubunit complexes containing both _________ & _________ activities; moves along the DNA in a reaction that requires ATP.

A

Endonuclease. Methylase.

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18
Q

Types of Cuts of Restriction E.: leaves 2-4 nucleotides of one strand unpaired at each resulting ends, can base pairs with each other or with complementary SE of the other DNA fragments.

A

Sticky ends

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19
Q

Types of Cuts of Restriction E.: cleaves both strands of DNA at opposing phosphodiester bonds leaving no unpaired base on the ends.

A

Blunt ends

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20
Q

Linear array of sites for the various restriction enzymes on a given piece of DNA.

A

Restriction map

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21
Q

Manual enzymatic method, Chain termination method and Dideoxy method.

A

Sanger

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22
Q

Chemical method

A

Maxam and Gilbert

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23
Q

The DNA being sequenced acts as a template. A complimentary chain is synthesized which elongates in 5’3’ direction. DNA chains of varying lengths are produced. The shortest chains are nearest the 5’ end of the DNA chain.

A

Sanger method

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24
Q

DNA to be sequenced is cleaved at specific bases by subjecting it to 4 diff. treatments in separate aliquots.

A

Maxam and Gilbert Method

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25
Q

M&G 4 Different Tx: reaction of DNA with ________ followed by piperidine treatment cleaves DNA before C and T.

A

C + T. Hydrazine.

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26
Q

M&G 4 Different Tx: If DNA is treated with hydrazine in __________, only C residues react appreciably; comparison of the C and C+T cleavage positions.

A

C only. 1.5M NaCL

27
Q

M&G 4 Different Tx: _________ methylates G residues at N7 rendering the glycosylic bond susceptible to hydrolysis; subsequent treatment with ________ cleaves the chain.

A

G only. Dimethyl Sulfate. Piperidine.

28
Q

M&G 4 Different Tx: Both A & G released when DNA is treated with ____________ causes strand cleavage before A & G residues; the A reidues are identified by comparing the position of G and A + G cleavages.

A

A+G. Acid Piperidine.

29
Q

DNA to be sequence is radioactively labeled with 32p at 5’ end; will produce a set of radioactive fragments whose members extend from 32p labeled end to one of the positions occupied by chosen base; sequence of DNA can then be directly read off on autoradiogram.

A

Maxam and Gilbert Method

30
Q

Large population of identical molecules, bacteria or cells taht arise from a common ancestor.

A

Clone

31
Q

Involves cutting a specific gene or DNA segment from a larger chromosome, attaching it to a small molecule of carrier DNA, and then replicating this DNA thousands or million of times through both an increase in cell number and the creation of multiple copies of the cloned DNA in each cell.

A

DNA Cloning

32
Q

In DNA Cloning: cutting DNA at precise locations with the use of restriction

A

Endonucleases

33
Q

In DNA Cloning: links the cloning vector and the DNA to be cloned. Composite of DNA molecules comprising covalently linked segments from two or more sources are called __________.

A

DNA Ligase. Recombinant DNA.

34
Q

A molecule into which foreign DNA can be introduced.

A

Cloning vectors

35
Q

Essential properties of Cloning Vectors: it must be capable of _______ replication within a host cell. It must contain atleast one specific _____________ recognized by a restriction endonuclease.

A

Autonomous. Nucleotide sequence.

36
Q

Essential properties of Cloning Vectors: It must carry at least ______ that confers the ability to select for the vector such as an antibiotic resistance gene.

A

One gene

37
Q

Small circular duplex DNA molecules whose natural function is to confer antibiotic resistance to the host cell. Exits as singke or multiple copies within the bacterium and replicate independently from bacterial DNA. Is known to precise location of restriction enzyme cleavage sites for inserting foreign DNA is available.

A

Plasmids

38
Q

Viruses that infect a bacterium. Have a linear DNA molecules into which foreign a DNA can be inserted at several restriction enzyme sites. It can accept DNA fragments 10-20 kb long.

A

Phages

39
Q

Plasmids that contain DNA sequences (called _____) required for packaging lambda DNA in the phase particle; unnecessary lambda DNA has been removed to chimeric DNA can be packaged into the particle head. It can carry DNA inserts 35-50 kb long.

A

Cosmids. Cos sites.

40
Q

Plasmids designed for cloning very long segments, 50-250 kb. Generally contain selectable markers such as resistance to the antibiotic ________. A very stable origin of replication (ori) that maintains the plasmid at 1-2 copies per cell.

A

Artificial chromosome

41
Q

Plasmids constructed for yeast since entire sequence of ___________ is known. It can carry DNA inserts of 500-3000 kb. Easy to maintain and grow on large scale in the laboratory.

A

Yeast Artificial Chromosome. Saccharomyces cerevisae.

42
Q

Vector in which the protein coded by the gene introduced by recombinant DNA is actually synthesized. Are constructed to contain very active inducible promoters, proper in phase translation, initiation codons, transcription and translation signals and appropriate protein processing signals if needed.

A

Expression vector

43
Q

Preparation of Chimeric DNA: technically easy. Disadvantage is that it may reconnect with themselves. Us of blunt ends with addition of new homopolymer tailing by a _________.

A

Sticky end ligation. Terminal transferase.

44
Q

Collection of cloned fragments that represents the entire genome.

A

Collection of cloned fragments

45
Q

Prepared from the total DNA of a cell line or tissues. Both introns and exons represented.

A

Genomic library

46
Q

Made from reverse transcription of the mRNAs in a tissue; only exons are represented.

A

cDNA library

47
Q

Is first digested with restriction endonucleases.

A

Genomic DNA or cDNA

48
Q

An in-vitro method of replicating relatively small DNA sequences into millions of copies over a short period. For rapid diagnosis of infectious diseases. In forensics, DNA from a single hair or sperm can detect the donor. To make prenatal/preimplantation genetic diagnosis. To study evolution using DNA from archaeological samples.

A

Polymerase Chain Reaction

49
Q

Short DNA sequences of approx 20bp in length designed to be complementary to the DNA sequences that flank of the DNA fragment of interest.

A

PCR primer

50
Q

Entails the localization or mapping of genes to each of 23 chromosome pairs. Determination of the order and spacing if the gene on the particular chromosome.

A

Gene mapping

51
Q

Based one estimates of physical distance measured in bp.

A

Physical mapping

52
Q

Method of chromosome visualization with the chromosomes identified by size and by distinctive banding patterns seen under light microscopy after staining with __________.

A

Karyotyping. Giemsa.

53
Q

Fulorescent tagged probes are prepared and used to localized the gene of interest to a particular chromosome.

A

FISH

54
Q

Based on the frequency of meiotic crossing over that is observed between 2 loci of a chromosome.

A

Genetic Linkage Mapping

55
Q

Variation in the sequence of a specific point in the DNA. May be revealed through RFLP and VNTR by __________.

A

Single Nucleotide Polymorphism. DNA Fingerprinting.

56
Q

Use of RFLPs or VNTRs to establish a unique pattern of DNA fragments for an individual. Used extensively in forensics and in paternity testing.

A

DNA Fingerprinting

57
Q

Variations in the length of restriction fragments resulting from sequence changes that creates or destroys a restriction site. I herited in the manner predicted by classical genetics.

A

Restriction Fragment Length Polymorphism (RFLP)

58
Q

Short DNA sequences that are repeated in sequence multiple times. Can be inherited. Useful in establishing genetic association with a disease in the family or kindred. Unique to an individual.

A

Variable Number of Tandem Repeats (VNTR)

59
Q

Variations in the short (2-6bp) inherited tandem repeat units of DNA that occurs about 50,000-100,000 times in the human genome.

A

Microsatellite DNA Polymorphism

60
Q

Small piece of silicon glass to which a number synthetic, single stranded DNA oligonucleotides probes have been chemically bonded. Used to monitor the expression of entire genome.

A

DNA MicroArray Chip

61
Q

Introduction of a gene for a missing or defective protein whose lack causes a disease

A

Gene Therapy

62
Q

Inactivation of a given gene which in turn creates strains of animals that lack the protein product of the gene,

A

Creation of knock out mice

63
Q

Introduction of mutations at specific points in a DNA sequence. Used to study the effect of changing single selected AA residue in protein. A chemically synthesized oligonucleotide is annealed to the DNA of the gene to be altered with the oligonucleotide as the primer.

A

Site directed mutagenesis

64
Q

Multicellular organism expressing a foreign gene.

A

Transgenic organisms

Biochemistry (20 decks)

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