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Frog

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by

Siqin Liu

on 17 October 2014

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Transcript of Frog

Frog
skin secretion/ skin fragment
mRNA
cDNA Library
precursor peptide-encoding cDNA
shot gun cloning
Database & BLAST
Putative sequence of mature peptide
the putative peptide would not be endogenous
secretion/fraction
HPLC
MS
nothing with same weight
compound with same weight
molecular weight compare
MS/MS
the putative peptide
is endogenous
'post-translational structure modification'
exists in the mature peptide
input
output
Verification
Sequence of Mature peptide
SPPS
Artificial mature peptide
Function test
Genetic information
Mature peptides
Separation
HPLC
Functional screening
Structure determination
Peptide sequence
Strategy Ⅰ
Strategy Ⅱ
Encoding cDNA sequence
Verification
NOTE: ' ' indicates the bio-information above would be added into NCBI database.
Amphibians
Peptide
Database
System support
Filling into
Hunting
Putative sequence
Sequence alignment
skin secretion/ fragment
Database
imput
What are we doing in this lab?
the hell
Our Research
到现在
范德萨
Man-made
Verification Procedure
Molecular cloning
Study on what?
What are the literatures talking about?
What are we doing now?
What is happening during the molecular cloning procedure?
How can the target gene be picked up?
mRNA isolation
cDNA library construction
Shotgun cloning--Polymerase Chain Reaction (PCR)
Gel analysis
Purification
mRNA:
DNA
cDNA
transcript
reverse transcript
What is complement DNA (cDNA)?
Diverse of cDNA sequences compose the Library
Primer:
All the target genetic information has been hunted .
Ligation & Transfermation
Blue & White Screening
Vector--Plasmid:
Exogenous DNA rings;
Transporter;
Selectable marker gene;
Multiple cloning sites;
Incompatiblity
JM109
Engineering bacteria
E.coli
Cellular proliferation
Cloning PCR
Gel-analysis
Purification
Sequencing
......
Sanger Sequencing
Correlation techniques & mechanisms
Blue & White Screening
Cell Culture
Polymerase Chain Reaction (PCR)
Rapid Amplification of cDNA Ends (RACE) PCR
Sangor Sequencing
Data mining & Sequence alignment in BLAST
Big number of new mechanisms should be dig out......
Digging technique which family strong?
Antimicrobial
Pharmacology
Anti-tumor
Synergistic effect
enzyme like/inhibitor
Amino acid sequence
Encoding DNA sequence
Submit the sequence to database
Searching
Alignment
Prediction
Full transcript