QTL Analysis of the OLETF Rat

    Quantitative Traits Loci (QTL) analysis of the animal model is one of the efficient approaches to identify the genetic determinants of complex human diseases, especially genes for polygenic diseases such as diabetes, hypertension, asthma, and cancer. To identify the genetic components associated with the diabetes and obesity, we took advantage of genetic dissection of the OLETF rat, obese type II diabetic model well representing human type II diabetes.
    For the analysis of diabetes and obesity-related phenotypes, we have used body weight, abdominal fat weight, plasma glucose (at fasting and during oral glucose tolerance test), plasma insulin, plasma triglyceride, plasma total cholesterol, and free fatty acid levels as indices. More than 200 markers are used for genetically-dissecting each cross, covering over 1,800 cM of the rat genome (supposed to be completely covered) with an average spacing of 8 cM or less (enough density to find out the QTLs with moderate effect). The comprehensive information of genetic linkage map and QTL analysis are viewed in the following section.

Crosses for genome-wide QTL analysis

    Note: Collaborative Linkage Analysis of Your New Animal Models Are Welcome: We have developed high-throughput genotyping system for extensive linkage analysis as well as unpublished genomic resources. Collaborative efforts to genetically dissect your animal model are considered. Please contact Takeshi K. Watanabe & Akira Tanigami for more information. Thank you.

Experimental Conditions

    The PCR was performed to assay all markers. Each 20ul reaction volume contained 50 mM KCl; 1.5 mM MgCl2; 10 mM Tris-HCl (pH 9.0); 5 % DMSO, 0.25mM of each dNTP, 20 ng of genomic DNA, 0.14 uM of each primer and 1 U of Taq polymerase (Amersham Pharmacia Biotech.). The PCR program was as follows: initial denaturation. 95 C for 3 min, followed by 40 cycles of 95 C for 1 min, 55 C or 52 C for 1 min and 72 C for 30 sec, followed by a final extension 72 C for 3 min. PCR products were applied on 10.0 % polyacrylamide mini-slab gels [90mm(W)x73mm(H)] (NPU-G10L-PAGEL, ATTO corporation, Japan) containing 25 mM Tris, and 192mM Glycine. Electrophoresis was run at constant current of 20 mA/gel for 75 min, and migrated DNAs were detected by silver staining.

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