what i’ve done this summer institute of molecular biology – academia sinica dr. che-kun james...
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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
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