Quantitative analysis of various PCDH11 X/Y gene transcripts - PowerPoint PPT Presentation

Title: Quantitative analysis of various PCDH11 X/Y gene transcripts


1
Quantitative analysis of various PCDH11 X/Y gene
transcripts
Kung Ahn1, Jae-Won Huh1, Dae-Soo Kim2, and
Heui-Soo Kim1,2
1Division of Biological Sciences, College of
Natural Sciences, Pusan National
University 2PBBRC, Interdisciplinary Research
Program of Bioinformatics, College of Natural
Sciences, Pusan National University
http//www.primate.or.kr
Protocadherins are members of the cadherin
superfamily that involved in cell-cell
interactions in the development of the central
nervous system. PCDHX/Y genes were known to be
composed of at least six exons sharing 98.1
identity. PCDHY is located on the Y chromosome
and originates from a translocation of the PCDHX
gene after the divergence between chimpanzees and
humans PCDH11 genes. All of PCDH11 gene related
transcripts were collected and analyzed using
bioinformatic analysis. From the EST and mRNA
sequence analysis, we could reconstruct the seven
different alternative transcripts of PCDH11X.
Specific elements of insertion and deletion were
identified by the multiple alignment analysis.
Those elements were used for the amplification of
individual transcripts. Specific amplification of
individual primer sequences was confirmed by the
analysis of melting curve. Absolute
quantification real-time RT-PCR amplification was
applied to compare seven different transcript
level of PCDH11X gene. One of the transcripts
shows the dominant expression pattern which seems
to be the indication of major roles among the
seven different transcripts. Taken together, we
suggest that the application of real-time RT-PCR
approach could be an advantageous method for the
investigation of closely related alternative
transcripts.
PCDHY is located on the Y chromosome and
originates from a translocation of the PCDHX gene
after the divergence between chimpanzees and
humans
Transcripts structures of the PCDHX/Y
x
Evolutionary changes in the genes transcribed
from the X/Y homology region in humans were
proposed to be causally linked to language
acquisition and functional brain asymmetry, two
hominid-specific features that are absent from
the brains of even our closest relatives,
chimpanzees.
translocation
Y
Standard curve and melt curve
in AJ276804
Materials Methods
Experimental Procedure
Primer Design
Twenty Different Human tissue cDNAs
Quantitative Analysis
Real-time RT PCR using SYBR Green I
Extension(I) phase
Annealing phase
One of the transcripts shows the dominant
expression pattern which seems to be the
indication of major roles among the seven
different transcripts.
SYBR Green I
Extension(II) phase
End of PCR cycle
1 Wilson ND, Ross LJN, Crow TJ, Volpi EV
PCDH11 is X/Y homologous in Homo sapiens but
not in Gorilla gorilla and Pan troglodytes.


2 Stuart N. Peirson,
Jason N. Butler and Russell G. Foster
Experimental validation of novel and conventional
approaches to quantitative real-time PCR data
analysis.
Detection of fluorescence
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