Cloning, Expression And Association Analysis Of The Marker Genes Of The Five Types Of Lymphocyte Located In The Immune Microenvironment Of The Porcine Endometrium And Involved In Immunological Tolerance

The modern Immunology of Reproduction believed that the embryo who regards as half homotransplant by the maternal immunity system is’t rejected is a very complex regulating process. The immunologic tolerance in this process is very important. The balance in matrix-embryo is affected by several factors. First, there are several kinds of immunocyte take part in immunodepression. Second, ratio of T cell subset is out of orderin the peripheral blood and endometrium. Third, the macrophage secrete kinds of cytokine, fouth, the activity of cytotoxic T cell (CTL) has fallen, fifth, the activity of NK cell has fallen. In this process various immunocyte play the most important roles.Each kind of immune cells have il’s special Marker. The Marker for Thl is transcription factor T-bet. Th2 is GATA3. FOXP3 transcription factor is for Treg. The Marker for Th17 is RORc. The macrophage take the leucocyte antigen CD 14 as its Maker. This research obtains the main results include:1. Has obtain the sequence of T-bet and RORc gene by electronic clone technology. Has predicted the amino acid sequence for the two genes by DNAMAN. Has annlyzed the protein struture information.2. Has screened pig the SNP sit for CD14, GATA3 and RORc. Has get their polymorphic site. These loci are as follows:g.844 A>G polymorphism in the porcine CD14 gene, g.1629 C>T polymorphism in the porcine GATA3 gene and g.1706 C>T polymorphism in the porcine RORc gene.3. PCR-RFLP was performed to detect the relations between polymorphism and its association with reproductive traits. Results indicated that the litter weight at birth of 4th parity is significantly (P=0.043).4. Association analyses revealed the significant associations (P<0.05) between the porcine GATA3 polymorphism and PLT measured on 0-day-old pigs and LYM, LY, MCH, PLT measured at 32 days after birth. The porcine RORc polymorphism was significantly associated (P<0.05) with RBC, HBG, P-LCR measured on 0-day-old pigs, respectively.5. Q-PCR was performed to detect the quantities of gene GATA3 in endometrium. Results indicated that the quantities is significant in the metaphase of pregnancy(p<0.05).