The Quantitative Detection Of FcRn In Gastrointestinal Organization And Its Sequence Analysis Of Piglets

In order to explore FcRn born - after weaning piglets in gastrointestinal organization, the expression and the correlation piglet intestinal IgG, taking hybrid binary piglets out as the research object,that sequence of gene was cloned and amplified and using different PCR RT - day-old binary piglets gastrointestinal tract FcRn level carries out quantitative variation.The main results were shown below:1.Total RNA was extracted from mucosal tissue of duodenum and mRNA sequence of gene were amplified by RT-PCR using two designed primers. The PCR products were ligated into the pMD-19T vector, and then transformed into competent cells of JM109. The sequence was analyzed to identify the recombinant plasmid. Identity analysis showed that the two FcRn nucleotide sequence shared 99 % .The porcine alternatively spliced variant of FcRn gene was successfully cloned in the present study and the sequence of spliced variant has been submitted to GenBank(Accession. EU852582).2.The results showed a significant difference of FcRn levels in swine tissues. The quantitation of FcRn levels in pgilet GI tract tissues was performed by quantitative real-time RT-PCR (RT-FQ-PCR) with primers and a fluorecent probe for FcRn heavy chain gene, withβ-actin chosen as house-keeping gene. The most abundance of FcRn was present in liver (△Ct =4.95), followed by distal duodenum and adjacent jejunum, and then adjacent duodenum and distal jejunum, low expression levels were revealed in ileum and colon, whilst the lowest level was in stomach (△Ct =6.91). The trend of FcRn mRNA expression in GI tract was present accordly to age: Low level of FcRn was foound at birth with no significant difference between non-suckling (0 w) and suckling (0 y) piglets (P>0.05), the expression markedly increased on 1 d (P<0.05), remained relatively stable and decreased at 21d, thereafter it increased sharply on 28d but dropped to previous level at 35d of age.