地高辛标记探针检测重组定点突变巴曲酶蛋白宿主DNA残留量的研究

Analysis of residual DNA in recombinant mutant batroxobin by using digoxin labeled probe

目的建立检测重组定点突变巴曲酶原液中外源性DNA残留量的方法。方法从重组定点突变巴曲酶酵母工程菌提取基因组DNA作为模板,制备地高辛标记探针。此探针与样品进行点样杂交,并进行显色反应。结果 3批重组定点突变巴曲酶原液中,每人份剂量的宿主DNA残留量均<10ng。结论该方法检测灵敏度较好,特异性较强,可用于重组定点突变巴曲酶的检测。

Objective To establish a method for testing the residual DNA in recombinant mutant batroxobin. Methods Genome DNA of engineering pichia pastoris of recombinant mutant batroxobin was used as template and as a probe labeled by digoxin. The dot blotting of probe hybridize with sample and continued to do chromogenic ere- action. Results The residual DNA in three batches of bulk products is less than 10 ng per dosage. Conclusion This method is sensitive and specific, which could be used for the quality control of recombinant mutant batroxo- bin.