杂交法检测重组乙型肝炎疫苗（汉逊酵母）发酵产物HBsAg外源基因拷贝数

摘要

The same MOX promoter fragments of Hansenula yeast host genomic DNA and fermen- tation products genomic DNA are taken as specific probes. Then the specific probes with digoxi- genin labeled are hybridized and been color with host genome DNA fixed in the hybrid mem- brane and fermentation products genome DNA. The copy number size of foreign gene in fermen- tation product according to the color depth can be determined. The results show that the copy number of HBsAg foreign gene in fermentation product is not less than 30. The method has such advantages as better detecting sensitivity, stronger specificity and good safety. Therefore, it can be used in qualitative detection of copy number size of HBsAg foreign gene in fermentation prod- uct of recombinant Hansenula yeast.%以汉逊酵母宿主基因组DNA和发酵产物基因组DNA的相同MOX启动子的片断为特异性探针,用地高辛标记后,与固定在杂交膜上的宿主基因组DNA和发酵产物基因组DNA进行杂交并显色,根据显色深度来判定发酵产物中外源基因拷贝数的大小。结果表明,发酵产物中HBsAg外源基因拷贝数不小于30个。该方法检测灵敏度较好,特异性较强,操作较安全,可用于重组汉逊酵母发酵产物中HBsAg外源基因拷贝数定性检测。