Quantitative analysis of various PCDH11 X/Y gene transcriptsQuantitative 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 University2PBBRC, Interdisciplinary Research Program of Bioinformatics, College of Natural Sciences, Pusan National University

http://www.primate.or.kr

References

Genome Information Lab

Introduction

Primer Design

Twenty Different Human tissue cDNAs

QuantitativeAnalysis

Experimental Procedure

Real-time RT PCR using SYBR Green I

Materials & Methods

Extension(II) phase

Extension(I) phase

End of PCR cycle

Annealing phase

SYBR Green I

Detection of fluorescence

Results & Discussion

Transcripts structures of the PCDHX/Y

Standard curve and melt curve in AJ276804

Expression pattern in various tissues

AbstractProtocadherins 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 advantageousmethod for the investigation of closely related alternative transcripts.

One of the transcripts shows the dominant expression pattern which seems to be the indication of major roles among the seven different transcripts.

[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.

PCDHY is located on the Y chromosome and originates from a translocation of the PCDHX gene after the divergence between chimpanzees and humans

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.