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Preliminary Study on the Porcine EDG4, EDG7 and OTUB1 Genes

Author: DanTongLing
Tutor: GuoDingZong；LiKui
School: Huazhong Agricultural University
Course: Clinical Veterinary Medicine
Keywords: pig LPA OTUB1 chromosomal localization Subcellular localization expression
CLC: S828
Type: Master's thesis
Year: 2008
Downloads: 31
Quote: 0
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Abstract


Lysophosphatidic acid （LPA） is a simple bioactive phospholipids with diverse physiological actions on many cell types, which secreted by platelet, fibroblast, malignant cell and cellula adiposa is a multipurpose "phospholipids messenger ".LPA plays an important role in the development of many kinds of momentous diseases, especially in cardiovascular disease, kidney disease and tumor by a. subgroup of G protein-coupled, endothelial differentiation gene （Edg）. EDG4 and EDG7 are the receptors of LPA, which transmit cellular message of LPA with Gi/o, Gq and G12/13, and play a key role in stimulus the growth of ovarian tumor. 0TUB1 gene is a member of the OTU （ovarian tumor） super family, which interacts with another ubiquitin protease that inhibits cytokine gene transcription in the immune system. The mechanism of action above-mentioned three genes needs further research in the development of ovarian tumor. Meanwhile, mini-pig and human are very similar in anatomy, physiology and pathology. Mini-pig is considered as an important laboratory animal of human disease model. In this study, we firstly cloned the above-mentioned three genes from porcine ovaries, investigated the expression of the genes and a series of analysis of a SNP in EDG4, in order to provide some information for the further study. In addition, we also investigated the chromosome location of porcine EDG4 and EDG7genes, and the Subcellular localization of porcine OTUB1 gene. The main results are as follows:1) Using bioinformatics and comparative genomics, EDG4, EDG7 and OTUB1 genes were cloned, and preliminary analysis of m RNA sequences and amino acid sequences of the genes.2) Using the INRA-University of Minnesota 7000 rads radiation hybrid panel （IMpRH）, EDG4 and EDG7 were respective mapped chromosome 2 and 6.3) The expression of the genes was investigated by RT-PCR. The results indicated OTUB1 gene was expressed in many tissues, mainly in immune organ and lowlier in muscular tissue. EDG4 and EDG7 were lower in many tissues.4) The green fluorescent protein （GFP） fusion expression vector of porcine OTUB1 gene was constructed, and was translated into PK15. Using confocal microscopy （Nikon C1）, fluorescent signals in the mitochondria and nuclei were analyzed at different excitation wavelengths; the protein was located in nucleolus. 5) Using PCR-RFLP, a SNP in porcine EDG4 gene was analyzed in domestic and overseas pigs. The result indicated gene frequency was more different between domestic and overseas pigs.6) Association analysis of the SNP was done. The result indicated there was different between heterozygote and homozygote in carcass length and drip loss percentage.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Pig
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