The Amplifying Of The Full CDNA And Tissue Expression Of Mongolian Sheep β-defensin-1

We mainly study Mongolian sheepβ-defensin-1. Total RNA was extracted from the rumen epithelial tissue of Mongolian sheep and the cDNA encoding sBD-1 was amplified by the reverse transcription PCR( RT-PCR ). The results of cDNA sequencing examined that the sBD-1 cDNA contain a open reading frame(ORF) of 192 bases which encoded a 64 amino acid prepro-peptide and the prepro-peptide contained the β–defensin consensus sequence of six invariantly spaced cysteine residues. We adopt the technic of Rapid Amplication of cDNA Ends.The anchored-PCR-RACE strategy was used to amplify 3ˊend of cDNA and backward nested- PCR-RACE strategy was used to amplify 5ˊend of cDNA.Then,they were identified to encode the full cDNA sequence of cBD-1 In order to examine the organs that express possibly sBD-1 mRNA in a Mongolian sheep, We examined the expression of sBD-1 mRNA in different tissues of the Mongolian sheep by RT-PCR. The result indicates that sBD-1 mRNA was detected in the mucosa of throughout the digestive tract except rectum ( tongue, esophagus, rumen, abomasums, reticulum,omasum,duodenum, jejumum, ileum, colon ), trachea, uterus and spleen, but sBD-1 mRNA wasn't detected in the heart ,liver, lung, kidney, lympho node, bladder,ovarium,oviduct.