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A Simple and Accurate Genotype Analysis of the motor neuron degeneration 2 (mnd2) Mice: an Easy-to-Follow Guideline and Standard Protocol Applicable to Mutant Mouse Models.
Hyun-Ah Shin1, Goo-Young Kim1, Min-Kyung Nam1, Hui-Gwan Goo2, Seongman Kang2 and Hyangshuk Rhim1,*
1Department of Medical Life Sciences, College of Medicine, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul 137-701, Korea
2School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Republic of Korea
*Corresponding author
  Received : August 29, 2012
  Accepted : September 12, 2012
  Published : September 13, 2012
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Synopsis

The motor neuron degeneration 2 (mnd2) mice carry a point mutation of A to T nucleotide transversion at the serine 276 residue of high temperature requirement A2 (HtrA2), resulting in losses of an AluI restriction enzyme site (5'AGCT3') and the HtrA2 serine protease activity. Moreover, dysfunctions of HtrA2 are known to be intimately associated with the pathogenesis of neurodegenerative diseases, including Parkinson's disease. Thus, this mnd2 mouse is an invaluable model for understanding the physiological role of HtrA2 and its pathological role in neurodegenerative diseases. Nevertheless, many molecular and cellular biologists in this field have limited experience in working with mutant mouse models due to the necessity of acquired years of the special techniques and knowledges. Herein, using the mnd2 mouse model as an example, we describe easy-to-use standard protocols for web-based analyses of target genes, such as HtrA2, and a novel approach for simple and accurate PCR-AluI-RFLP-based genotype analysis of mnd2 mice. In addition, band resolution of AluI-RFLP fragments was improved in 12% polyacrylamide gel running in 1X Tris-Glycine SDS buffer. Our study indicates that this PCR-AluI-RFLP genotype analysis method can be easily applied by the molecular and cellular biologist to conduct biomedical science studies using the other mutant mouse models.

Keyword: mnd2 mice, HtrA2, genotyping, web-based analyses, PCR-RFLP, mutant mouse
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