PLANTBI C112 Chapter Notes - Chapter 12: Mutagen, Yazh, Green Fluorescent Protein
19 Sep 2018
School
Department
Course
Professor
Ch .12 :Biotech
12.I.
genetic engineering :use of in vitro techniques to alter genes in the lab
.heterologous expression
:exp . of agene from 1org in adiff .host
.PCR
.
.DNA rept. in vitro
,amplification
-need primers made of DNA
,
not RNA
-usually 2030 cycles
-Tag polymerase .
.from hyperthermophilic bait
-Pfu polymerase :proofreads ,
for high accuracy
-Ssu rRNA .
.
conserved
.
for phylog.
-
reverse transcription PCR (RT -Pek):make DNA from mRNA template ,
to detect expression
-
reverse transcriptase : in RT .PCR
,
RNA → cDNA
,
results in hybrid 4DNA tRNA
-cDNA : complementary DNA
-RN an It :hydnliccs RNA
-hybridization :
complementary
,
for detention
-Southern blot :probes hybridized to target DNA fags that have beenSep .by electrophoresis
-Northern blot : uses RNA as target sequence ,to detect gone expression,gives rough
est . of mRNA abmd . of target gun
II.
2.vector :manipulable genetic
element .
i.
molecular cloning:
movement of desired genes from orig source to veto r
recombinant DNA :DNA from diff .sources
.palindromes :sq .that nest . en z.
recognize
.
sticky ends :short
,
sis . overhangs
.blunt ends .
.
after rest . eat cuts
,nothing hanging off
.DNA ligase :joins sticky ends
.multiple cloning sik :short segment w/ multiple wt sites
.insertion inactivation :after foreign DNA added to numb . vector
,
.
yeast attic ial chromosomes
.(YAL s):for cloning genes into yeast,
have origin of DNA replication
2. 3obstacles to gene exp r
.
of cut .some
-
genes must be placed under
control of bad .promoter
-introns must be removed
Document Summary
, amplification use of in vitro techniques to alter genes of a gene host from 1 org in a diff the lab in. Need primers made of dna not rna. Pfu polymerase : cycles from hyperthermophilic bait proofreads , for high accuracy. Ssu reverse reverse rrna . conserved for phylog . transcription pcr (rt - pek) : make dna from mrna template to detect expression transcriptase : in rt . Rna cdna results in hybrid 4 dna trna. Southern blot : probes hybridized to target dna fags that have been. Northern blot : est uses rna as target sequence , of mrna abmd . of target gun. 2 . vector : manipulable genetic element . to detect gone expression. 2 guns short recognized seg . between signal sequence. : peptide rich in hydrophobic aa , that enables transport. 4 site - directed1in vitro mutagenesis uses synth dna cloning techniques to intnd .
