畜牧兽医学报2012，Vol.43Issue(9)：1377-1384,8.
绵羊Gfrα1基因的部分cDNA克隆与抗原表位多肽的原核表达
Cloning of Gfrα1 Partial cDNA in Sheep and Prokaryotic Expression of Epitope Polypeptide
摘要
Abstract
In order to clone the DNA sequence of Gfral gene in sheep and deeply research the sheep spermatogonial stem cells (SSCs), in the present study, the partial sheep Gfral cDNA were cloned using RT-PCR and molecular cloning technique. The cloned partial Gfral gene was 1 312 bp in length, including an ORF of 1 311 bp encoding 437 amino acids. The nucleotide sequences of sheep Gfral gene were compared with the counterpart sequences of Bos Taurus , homo sapiens , Pongo abelii , Rattus norvegicus and Mus musculus , and the nucleotide homology was 98. 5% , 91. 3 % , 91.0%, 88. 9% and 88. 0% , respectively. According to the cloned Gfral gene sequence, the epitope was forecasted and then the cDNA was inserted into pET-44a( + ) vector. Subsequently, the recombinant was induced by IPTG. The recombinant epitope polypeptide was purified according to Affinity column chromatography. Western blot analysis indicated that the molecular weight of the expressed epitope polypeptide was the same as predicted size of approximate 18. 2 ku. The data collected provided the important information for further making Gfral gene polyclonal antibody and authenticating on molecular level for spermatogonial stem cells in sheep.关键词
绵羊/Gfrα1基因/克隆/序列分析/原核表达Key words
sheep/ Gfral gene/ cloning/ sequence analysis/ prokaryotic expression分类
农业科技引用本文复制引用
刘燕,罗奋华,包佳婧,刘林洪,单飞彪,吴应积..绵羊Gfrα1基因的部分cDNA克隆与抗原表位多肽的原核表达[J].畜牧兽医学报,2012,43(9):1377-1384,8.基金项目
教育部创新团队计划子课题(IRT0833) （IRT0833）
高等学校博士学科点博导类基金项目(20101501110001) （20101501110001）
