Plant Biotechnology Methods

Question 1

Organism containing foreign DNA

Answer 1

Transgenic

Question 2

Process of inserting DNA into genome of cells

Answer 2

Transformation

Question 3

Use of isotope type: 3H

Answer 3

Most biological molecules

Question 4

Use of isotope type: 14C

Answer 4

Most biological molecules

Question 5

Use of isotope type: 35S

Answer 5

Proteins

Question 6

Use of isotope type: 32P

Answer 6

Nucleic acids, Phosphoproteins

Question 7

Beta particle detector: Meter that counts current pulses and calculates rate (cps or cpm); typically used to survey for workplace contamination

Answer 7

Geiger-Mueller Detector

Question 8

Beta particle detector: radioactive particles or photons produce series of excited states in fluor, output of fluor proportional to energy of interacting particle or photon

Answer 8

Liquid scintillation counter (LSC)

Question 9

Small dsDNA with internal RE site, blunt-end ligated to cDNA, RE digestion of linker for insertion into vector

Answer 9

Linkers

Question 10

Linkers with preformed sticky ends

Answer 10

Adapters

Question 11

Fragment of DNA complementary to Gene of Interest

Answer 11

Gene probe

Question 12

Gene you have and want to see what interacts with it

Answer 12

Bait

Question 13

Protein that interacts with bait

Answer 13

Prey

Question 14

Homologous genes from another organism

Answer 14

Heterologous probes

Question 15

Requires knowledge of DNA or protein sequence

Answer 15

Synthetic DNA probes

Question 16

Requires knowledge of DNA sequence to design PCR primers

Answer 16

PCR

Question 17

Which method of DNA labeling involves Klenow fragment of DNA polymerase?

Answer 17

Random primer labeling

Question 18

A large protein fragment that is produced when DNAP1 from E. Coli is enzymatically cleaved, has lost 5’ to 3’ exonuclease activity

Answer 18

Klenow fragment

Question 19

Which method of DNA labeling involves DNAP1 and labeled dNTPs ?

Answer 19

Nick translation labeling

Question 20

Which method of DNA labeling involves attaching a reporter enzyme to DNA ?

Answer 20

Non-radioactive direct labeling

Question 21

Which method of DNA labeling involves specific binding proteins, nucleotides, and a reporter enzyme?

Answer 21

Non-radioactive indirect labeling

Question 22

Blotting technique used for DNA

Answer 22

Southern blotting

Question 23

Blotting technique used for RNA

Answer 23

Northern blotting

Question 24

Which cloning method of antibodies involves forming hydridomas ?

Answer 24

Monoclonal antibodies

Question 25

Which cloning method of antibodies involves drawing blood from the host and recovering the serum ?

Answer 25

Polyclonal antibodies

Question 26

A good primer for PCR is typically how long?

Answer 26

20-30 bp

Question 27

Maintain and propagate cloned DNA

Answer 27

Cloning vectors

Question 28

Four different types of vectors

Answer 28

Plasmid, Phage, Cosmid, Artificial chromosome

Question 29

Extrachromosomal DNA in bacterial cells; carries genes for antibiotic resistance, conjugation, and metabolism of unusual substrates

Answer 29

Plasmid

Question 30

ORI that replicates with cell cycle

Answer 30

Stringent ORI

Question 31

ORI that replicates faster than cell cycle

Answer 31

Relaxed ORI

Question 32

Hybrids of phage lambda and plasmids; infects cell like a phage, replicates like a plasmid

Answer 32

Cosmid

Question 33

Large insert capacity cloning vectors

Answer 33

Artificial chromosome

Question 34

Linear molecule, inserted into SUP4 (suppressor tRNA marker)

Answer 34

Yeast artificial chromosome

Question 35

DNA sequence with regulatory elements to get desired expression in the transformed plants

Answer 35

Expression cassette

Question 36

What are the components of an expression cassette ?

Answer 36

Gene of interest, promoter, protein localization signal (if necessary), 5'-UTR and 3'-UTR sequences, and a selectable marker

Question 37

What are the ways of obtaining a gene of interest ?

Answer 37

Genomic library screens, cDNA library screens, PCR, gift from someone who has gene

Question 38

What is the most commonly used constitutive primer?

Answer 38

35S Cauliflower Mosaic Virus promoter

Question 39

What is a chimeric promoter?

Answer 39

cis element from plant inducible genes + 35S promoter

Question 40

What are the 5 components of MS medium?

Answer 40

Major inorganic nutrients, minor inorganic nutrients, iron source, organic supplement, and carbon source

Question 41

Required specifically for root growth, more than one compound can behave like this one, also required for callus culture, somatic embryo induction, and plantlet regeneration

Answer 41

Auxin

Question 42

Required specifically for sheath growth, promotes cell division, also required for callus culture, sometimes somatic embryogenesis and plantlet regeneration

Answer 42

Cytokinins

Question 43

Large class of compounds of related structures, GA3 sometimes used to promote growth

Answer 43

Gibberellins

Question 44

Used to promote distinct developmental pathways

Answer 44

Abscisic acid

Question 45

Unorganized growing and dividing mass of cells

Answer 45

Callus

Question 46

Densely aggregated callus

Answer 46

Compact

Question 47

Loosely associated, easily broke apart callus

Answer 47

Friable

Question 48

Plant cells with cell walls removed

Answer 48

Protoplast