What I

1
What Ive done this summer

• Institute of Molecular Biology Academia Sinica
• Dr. Che-Kun James Shen ?????
• National Health Research Institutes
• Dr. Xin Chen ????
• 891619
  ???

2
Recent Projects of Dr. Shen

• Brain cDNA Library Sequencing
• Sequencing of macaques brain cDNAs
• Compare macaques brain cDNAs with humans.
• 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 gene expression quantity between right
  and left brains of mice.

3
My work in Dr. Shens Lab

• Learned some concepts of PCR
• Preliminary screening for target genes derived
  from right and left brains of mice
• Qualitative RT-PCR

4
Introductions to Glycophorin Gene Family

• 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.

5
Why Are We Interested in Glycophorin Gene Family ?

• 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.

6
Methods
PCR with glycophorin-specific primers
Blue-white selection
Gel electrophoresis
Colony PCR
Vector-insert ligation
Plasmid DNA sequencing
Transformation
Data analyses
7
Results
8
My work in Dr. Chens 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
9
My work in Dr. Chens 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