中国畜牧兽医2017，Vol.44Issue(3)：635-643,9.DOI:10.16431/j.cnki.1671-7236.2017.03.003
RACE法扩增梅花鹿β-防御素-1(siBD-1)全长cDNA
Amplification of the Full-length cDNA of Sika β-defensin-1(siBD-1) with RACE Method
摘要
Abstract
In order to obtain the full-length cDNA of sika β-defensin-1 (siBD-1),5′-RACE and 3′-RACE primers were designed according to the partial sequences of siBD-1 that obtained from this study group, cDNA ends (RACE) technology were used to amplify siBD-1 cDNA from the total RNA of tongue mucosa tissue in sika, and amplified products were cloned into pMD18-T vector and subjected to PCR, restriction endonuclease digestion and sequencing.The results indicated that the length of siBD-1 cDNA 5′-and 3′-end were about 172 and 299 bp.The full-length siBD-1 cDNA was 418 bp (GenBank accession No.HM588696.1) and includes an 5′-untranslated region (UTR) of 89 bp, a open reading frame (ORF) of 192 bp, a stop codon of TAA, 3′-UTR of 118 bp and Ploy(A)16.The sequence homology showed that siBD-1 shared the greatest identity (90.6%) with buffalo enteric β-defensin, the higher identity with cattle (EBD, LAP, TAP, BNBD-4), goats (GBD-1, GBD-2), reindeer (reBD-1), sheep (sBD-1, sBD-2) and camel (caBD-1), that were 83.2%, 83.1%, 87.3%, 87.0%, 87.5%, 87.5%, 84.4%, 79.9%, 77.1% and 70.5%,respectively,and correspondingly low identity were 60.3% and 72.4% with horse (hoBD-1), pig(pBD-1), the lowest was 16.0% with human (hBD-2).The mature peptide was consisted of 38 amino acids, and contained 9 positively charged residues.关键词
梅花鹿/5′-RACE/3′-RACE/β-防御素-1Key words
sika deer/5′-RACE/3′-RACE/β-defensin-1 (siBD-1)分类
农业科技引用本文复制引用
唐娟,金鑫,张曼,侯永跃,李军燕,田巧珍,刘骄,王云鹤,杨银凤..RACE法扩增梅花鹿β-防御素-1(siBD-1)全长cDNA[J].中国畜牧兽医,2017,44(3):635-643,9.基金项目
内蒙古自然科学基金(2013MS0408) （2013MS0408）
