The Aquaporins Gene MRNA Expression Diversity After Weaning In Difference Tissue Of Piglets Using SYBR Realtime PCR

Aquaporins (AQPs) was one kind of the membrane channel proteins which could transport water molecules selectively and efficiently. The study showed that AQPs expressed abundantly in intestinal epithelial cells, and AQPs was involved in the secretion of intestinal mucus. The piglet's diarrhea was the most common, harmful problem for the early weaned piglets, which induced with the super-early-weaned piglets (SEW) technology. Therefore, making the role of AQPs in the pathogenesis of weaned piglets diarrhea, appears very important, which could provide theoretical basis for the clinic treatment, improve treatment technology and means.In this paper, took pig as object of study, designed primers according to the cloning principle of homologous sequence, cloned pig Aqp2-11 gene by RT-PCR, and then run bioinformatics analysis. The expression level of nine housekeeping gene ACTB,B2M,GAPDH,HMBS,HPRT1,RPL4,SDHA,TBP1 and YWHAZ mRNA was detected in different tissues of piglets by real-time fluorescence quantitative PCR technology, identified the best gene to calibrate target gene expression in different tissues of piglets. Then selected the gene Aqp1, 3, 7, 8, 10 and 11 of pig, and the diversity of mRNA expression was detected by real-time fluorescence quantitative PCR before and after weaning, and the results showed that:1) The cloned AQPs gene family include Aqp2, Aqp3, Aqp4, Aqp5, Aqp6, Aqp7, Aqp8, Aqp9, Aqp10 and Aqp11, and the corresponding NCBI accession number are EU636238, EU161094, EU165525, EU192130, EU620575, EU024116, EU220426, EU220427, EU582021 and EU220425. Sequence analysis showed that: the selectable permeability of pig AQPs gene family comply with a standard of the four sub-family classification.2) The expression stability of the housekeeping gene was: HMBS and HPRT1> RPL4> TBP1> B2M1> YWHAZ> SDHA> GAPDH> ACTB, which identified that the HMBS gene was the best choice to calibrate target gene expression in different tissues of piglets, and RPL4 gene could be taken as the internal gene of the high transcripts. 3) After the piglet was weaned, the expression of pig AQP1, AQP3, AQP7, AQP8, AQP10 and AQP11 generally increase in the digestive tract. And the expression of AQP8 in the gastric tissue after weaning significantly increase.