The Coding Sequence Cloning And Expression Analysisi Of EphrinA1 And EphB2 During Embryo Implantion In Meishan Pigs

In order to study the reproductive traits of erythropoietin genes relating to produce liver cells(EphrinA1)and Eph B2 receptor ligands A1 receptors(EphB2)of meishan pig embryos with plant,the influence of the litter size,the following study have been carried out in this paper.1.The embryo with plant period of meishan pig cloning and expression EphrinA 1 all coding regions and bioinformatics analysis(Objective):In order to analyze the role of EphrinAl in the migration and adhesion of embryo along the endometrium during embryo implantation in Meishan swine.(Methods):Using the sample,EphrinAl coding sequence was cloned,protein structure and function were predicted and mRNA expression was detected using cloning,sequencing,bioinformatics and qRT-PCR method.(Results):The results showed that there were five cSNPs were detected,show the child out 1 has two mutations were G89A,G91C,show the child outside 2 also has two mutations were T101C,A208G,outside show is a mutation happened on 4 children G526A,and two cSNPs caused two animo acids change,including Glu29Asn in Exon 1 and Trp33Arg in Exon 2.EphrinA1 coding sequence contained five exons and encoded 205 amino acids,EphrinAl protein was hydrophilic and instable,has a signal peptide and a conservative region and no transmembrane region.Moreover,EphrinA1 mRNA had a strong expression in Meishan pigs during embryo implantation period.It can be inferred from that specific binding of protein may be caused by mutations of Glu29Asn and Trp33Arg,which would affect the function of EphrinAl gene.In addition,EphrinA1 plant period in meishan pig embryos with strong mRNA expression,EphrinA1 in embryonic attached plant period(13 to 24 days)mRNA expression reduced trend after the first increases,namely in the process of embryo with plant early to mid express volume continued to rise,in the process of embryo of mid to late graft expression quantity continues to reduce.And the expression of pregnancy day 18 quantity is significantly higher than other gestational age(P<0.05),the expression quantity EphrinAl mRNA expression in meishan pig embryos with plant regulation may play a role.whether the EphrinAl high mRNA expression quantity participate in the embryo implantation regulation of meishan pig is need to be analyzed at the cellular level through gene expression and excessive silent technology.(Conclusions):The data analyzed from meishan pigs EphrinA1 gene cloning and expression CDS area and biological showed that these may participate in the regulation of embryo implantation in meishan pig,and influence the function of EphrinAl gene.and new cSNPs is a potential molecular marker affecting swine litter size.2.Meishan pig embryonic organization of planting period EphB2 expression analysis(Objective):The liver cell B2 receptor(EphB2)produced by Erythropoietin play a role in a variety of cell migration and invasion activity.As a member of the family Eph-Ephrin receptor,EphB2 play a role in a variety of cell migration and adhesion work.A study reported EphB2 excessive expression can reduce human glioma cells adhesion ability and increase the invasive ability of the cell.In order to study the impact of EphB2 to the stage of embryo implantation of pig and the migration and adhesion of the endometrium.(Method):We selected the meishan pigs in taihu lake basin in as test object,using the method of real-time fluorescent quantitative PCR and Western blot method,analyzed the gene in meishan pig embryos with plant before,during and post endometrial and ovarian tissue mRNA and protein expression spectrum.(Results):Showed that mRNA expression of EphB2 stay the same in the stage of embryo implantation(13 to 24 days),which is rising before reducing.The expression quantity of 18 th days is significantly higher than other gestational age(P<0.01).In the ovary,mRNA expression of EphB2 in the stage of embryo implantation is reducing before rising and the expression quantity of 18 th days is significantly lower than other gestational age(P<0.01),which is opposite to it in endometrium.In addition,in the 18 th days of pregnancy,the expression of the endometrial attached to the plant was significantly higher than that of endometrial attached point between the graft and ovarian tissue(P<0.01).the protein expression curve of the EphB2 in Endometrium with points and with plant between points was reduced after the first rise trend,namely the expression quantity in the process of embryo implantation early to mid continued to rise,the expression quantity in the process of embryo implantation mid to late continued to reduce.Thus EphB2 in embryonic phase graft swine endometrial mRNA and protein expression are presented after the first rise to reduce the trend,and the attached plant medium of expression is significantly higher than early and later(P<0.01);The EphB2 expression in ovarian trend,on the other hand,after the first reduce rise.(Conclusion):EphB2 is likely play an important role in regulation in the process of pig embryo implantation,which can be a potential candidate genes of swine litter size traits.Above all,EphrinA1 and EphB2 gene in meishan pig embryonic period has a high expression of the quantity,may to the pig embryonic attached plant plays a certain regulation effect,can be a pig litter size traits candidate genes.