Clonging And Expressing Analysis Of OPN In Yorkshire

Osteopontin (OPN) is an secretion type glycosylation phosphoprotein, which contains anArg-Gly-Asp (RGD) sequence, and it is classified a component of the extracellular matrix (ECM).The subsequent research found that OPN was expressed in cellula epithelialis uterineendomembrane, the invaded trophoblastic cell, the glandular epithelium of decidua and placenta inthe secretion phase. Furthermore, OPN is expressed on the maternal-fetal interface continuouslyand inferred that OPN play a key role in the regulation of uterine-placenta microenvironment.Therefore, it has important significance for studying on pig's candidate genes of reproductive traitthat molecular genetic construction and space-time expression of OPN gene is studied by modernmolecular biological technology.This research applied to Yorkshire, wild boar and Crossbred pig as the object of studying,OPN gene of Yorkshire and the wild boar was cloned, and its structure function and the molecularevolution had analyzed, at the same time, expression situation of OPN gene of Yorkshire andCrossbred was analyzed in the heart, the liver, the stomach, the spleen, the kidney, the lung, thelarge intestine, the small intestine, the muscle, the uterus and the ovary. Main research results asfollows:Getting completed OPN cds of Yorkshire and wild boar. From sequence comparation we foundthat there are five mutant sites in Yorkshire and eight in wild boar. The analysis on domin of OPNindicate that there is only one characteristic domin in OPN. Phyletic anlaysis found that OPN of pighas high homology with cattle, bufflo,and sheep but low with fowl.The results of RT-PCR indicate that OPN of Yorkshire expressed in eleven tissues at diffentdevelopmental stage, and did not have obviously regulatory. In the glandular tissue, OPN expressedincreasingly as time passed by. The results of semi-quantitative RT-PCR indicate that OPNexpressed differently in Yorkshire and wild family hybrid pig. But both in Yorkshire and wildfamily hybrid pig the expression trend is high to low to high and reach the peak. The peak ofYorkshire is in the 180 days and wild family hybrid pig is 360 days.The results of Western blot indicate that OPN expressed differently in different tissues. OPNmotified differently in different tissues, which lead to different hydrolization degree and diversityexpression format.Induced expressing and RT-PCR detecting OPN expreesion variation in endometrium endothelial cell with different hormones. The detected results found that pregnancy hormone increased OPNexpressing but hormones decreased.