What I’ve done this summerWhat I’ve done this summer

Institute of Molecular Biology – Academia SinicaInstitute of Molecular Biology – Academia SinicaDr. Che-Kun James Shen Dr. Che-Kun James Shen 沈哲鯤博士沈哲鯤博士

National Health Research InstitutesNational Health Research InstitutesDr. Xin Chen Dr. Xin Chen 陳新博士陳新博士

891619 891619 陳惠民陳惠民

Recent Projects of Dr. Shen Recent Projects of Dr. Shen

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.

Brain asymmetrical gene expression– Find some target genes which have notable different ge

ne expression quantity between right and left brains of mice.

My work in Dr. Shen’s LabMy work in Dr. Shen’s Lab

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PCR cycle數

DN

A量 X左腦 基因

X右腦 基因 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 FamilyIntroductions to Glycophorin Gene Family

GPA, GPB and GPE are highly homologous and form GPA, GPB and GPE are highly homologous and form a gene cluster on chromosome 4a gene cluster on chromosome 4 （（ q28 - 31q28 - 31）） ..

The antigens for the MNS blood group system are The antigens for the MNS blood group system are GPA and GPB.GPA and GPB.

The existence of about 40 variant phenotypes of this The existence of about 40 variant phenotypes of this blood group system has been documented by blood group system has been documented by serological analyses.serological analyses.

Why Are We Interested in Glycophorin Gene FamiWhy Are We Interested in Glycophorin Gene Family ?ly ?

The allelic diversity arises from The allelic diversity arises from unequal homologous crossing-over or unequal homologous crossing-over or gene conversions rather than point gene conversions rather than point mutations.mutations.

The incidence of the allelic diversity The incidence of the allelic diversity across the world appears to be across the world appears to be characteristic of the ethnic or characteristic of the ethnic or geographic origin of the subjects.geographic origin of the subjects.

The evolution of the three identified The evolution of the three identified hot spots.hot spots.

Blood group antigen have become Blood group antigen have become classic genetic markers in genetic classic genetic markers in genetic population studies and in linkage population studies and in linkage analyses.analyses.

MethodsMethods

PCR with glycophorin-specific primers

Gel electrophoresis

Vector-insert ligation

Transformation

Blue-white selection

Colony PCR

Plasmid DNA sequencing

Data analyses

ResultsResults

My work in Dr. Chen’s LabMy work in Dr. Chen’s Lab

To learn how to determine the optimal condition for protein To learn how to determine the optimal condition for protein purification.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 LabMy work in Dr. Chen’s Lab （（ ContinuedContinued ）） 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