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卵清蛋白基因表达放射免疫分析法
引用本文:朱献玳,王公金,严建民,张德玉.卵清蛋白基因表达放射免疫分析法[J].核农学报,1993,7(1):52-56.
作者姓名:朱献玳  王公金  严建民  张德玉
作者单位:江苏省农业科学院原子能农业利用研究所,江苏省农业科学院原子能农业利用研究所,江苏省农业科学院原子能农业利用研究所,江苏省农业科学院农业生物遗传生理研究所 南京 210014 ,南京 210014 ,南京 210014 ,南京 210014
摘    要:应用竞争结合原理建立了卵清蛋白基因表达的放射免疫分析法。采用氯胺T法标记卵清蛋白,~(125)Ⅰ-OA比活度达32~43μCi·μg~(-1),冻干后在常温下保存,有效期2~3个月;以(鸡)卵清蛋白作免疫原制备出的抗血清特异性强,与其他生物大分子无交叉反应,亲和常数为3.8×10~9L/M,50%结合率为1:1.6×10~5(最终稀释度);用双抗-PEG作分离剂沉淀免疫复合物,其方法灵敏度为0.17ng,可检范围为0.25~16ng,批内、批间变异系数分别为2.99%和8.96%。该方法能有效地直接检测类卵清蛋白含量,可作为一种特异、灵敏、简便、可靠的分析技术,应用于卵清蛋白基因表达生物基因工程研究。

关 键 词:卵清蛋白    基因表达    放射免疫分析
收稿时间:2009-12-31
修稿时间:2009-12-31

A RADIOIMMUNOASSAY METHOD FOR DETERMINATION OF OVALBUMIN GENE EXPRESSION
Abstract:A radioimmunoassay method for determination of ovalbumin-like protein was established on the principle of competition binding assay.The antigen ovalbumin (chicken) was labelled with 125I by Ch-T method.The specific activity of the iodinated antigen was 32-43μCi/μg,it was freeze-drying,stored under room tempreture and could be used for about three months.The anti-serum against ovalbumin (chicken )had no cross-reaction with the other biomacro-molecules.The affinity constant (K) of antiserum against ovalbumin-like protein was as high as 3.81×109L/M,the final dilution of antiserum was 1 : 160000.In this procedure,second antibody-PEG would be used as a precipitant to isolate the immune compound.The sensitivity was 0.17ng and the range of the determination for samples was 0.25-16 ng.The coefficient variation of intrabatch and in-terbatch were 2.99% and 8.96 % respectively.The method could successfully and directly determine the concentration of ovalbumin-like protein in samples.The radioimmunoassay method described here is specific,sensitive,reliable,simple and convenient for studying gene expression of ovalbumin-like protein.
Keywords:ovalbumin-like protein  gene expression  radioimmunoassay
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