Transcript What I`ve done this summer

What I’ve done this summer
Institute of Molecular Biology – Academia Sinica
Dr. Che-Kun James Shen 沈哲鯤博士
National Health Research Institutes
Dr. Xin Chen 陳新博士
891619 陳惠民
Recent Projects of Dr. Shen
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Brain cDNA Library Sequencing
– Sequencing of macaque’s brain cDNAs
– Compare macaque’s brain cDNAs with human’s.
– Expect to find some candidate genes which cause the
“superiority” of humen over other primates.
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Brain asymmetrical gene expression
– Find some target genes which have notable different
gene expression quantity between right and left brains
of mice.
My work in Dr. Shen’s Lab
20
DNA量
15
左腦X基因
10
右腦X基因
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5
0
0
10
20
30
40
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PCR cycle數
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Learned some
concepts of PCR
Preliminary screening
for target genes
derived from right
and left brains of
mice
Qualitative RT-PCR
Introductions to Glycophorin Gene Family
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GPA, GPB and GPE are highly homologous and form
a gene cluster on chromosome 4（q28 - 31）.
The antigens for the MNS blood group system are
GPA and GPB.
The existence of about 40 variant phenotypes of this
blood group system has been documented by
serological analyses.
Why Are We Interested in Glycophorin Gene
Family ?
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The allelic diversity arises from
unequal homologous crossing-over or
gene conversions rather than point
mutations.
The incidence of the allelic diversity
across the world appears to be
characteristic of the ethnic or
geographic origin of the subjects.
The evolution of the three identified
hot spots.
Blood group antigen have become
classic genetic markers in genetic
population studies and in linkage
analyses.
Methods
PCR with glycophorin-specific primers
Blue-white selection
Gel electrophoresis
Colony PCR
Vector-insert ligation
Plasmid DNA sequencing
Transformation
Data analyses
Results
My work in Dr. Chen’s Lab

To learn how to determine the optimal condition for protein
purification.
Inoculation and incubation overnitht
Induction（Different Time, oC and [IPTG]）
Ultrasonication
centrifucation
SDS gel electrophoresis
comassive blue staining
My work in Dr. Chen’s Lab（Continued）

Plasmid construct for future work
PCR for the target gene
Use restriction enzymes to treat both inserts and vectors.
（Check the frame in advance.）
Ligation（Check the insert-vector ratio.）
Transformation
Use the same Restriction enzymes
to check whether the insert is inside or not.
Plasmid DNA sequencing