地高辛标记核酸探针检测重组人白细胞介素1受体颉颃剂中DNA残余量的研究  

Detection of Residual DNA in Recombinant Interleukin-1 Receptor Antagonist(IL-1ra) Using Digoxigenin Labeled Probe

作　　者：王超[1] 刘庚[2] 张林波[1] 

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118 [2]吉林农业大学中药材学院,吉林长春130118

出　　处：《中国畜牧兽医》2013年第11期54-58,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然基金(31072127)

摘　　要：试验旨在建立重组人白细胞介素1受体颉颃剂(interleukin-1receptor antagonist,IL-1ra)原液中宿主菌DNA残余量的检测方法。以重组白细胞介素1受体颉颃剂工程菌基因组DNA为模板,优化试验条件,借助超声波手段,制备地高辛标记的核酸探针,并以此探针进行狭缝杂交及显色反应。结果显示,在24批次重组人IL-1ra原液中,每人份使用剂量的宿主DNA残余量均小于10ng。结果表明,该方法灵敏度较高、特异性较强、重复性较好、操作步骤安全且较为方便,可用于重组人IL-1ra原液及半成品的DNA残余检测。The assay was aimed to establish a method for detecting the residual DNA in recombinant interleukin-1 receptor antagonist （IL-lra） using digoxigenin labeled probe. By optimizing experimental conditions, with ultrasonic means,genomic DNA of engineered bacteria containing IL-lra was labeled by digoxigenin,as a probe for slot blotting assay to dectect residual DNA in bulk products. The residual DNA in 24 batches of bulk products was less than 10 ng per dosage for per person. This method was sensitive, specific and convenient,and could be well repeated, which could be used for detecting residual DNA of recombinant IL-lra protein bulk products and semi-finished products.

关 键 词：DNA残余量 地高辛 重组人白细胞介素1受体颉颃剂 狭缝杂交 

分 类 号：Q78[生物学—分子生物学]