摘要
【目的】探讨采用荧光定量PCR技术检测脑源性神经营养因子 (BDNF)基因转染细胞 (PcDNA3 1(+) /BDNF/CHO)中BDNF的拷贝数。【方法】分别以同等质量的PcDNA3 1(+) /BDNF/CHO ,PcDNA3 1(+) /CHO(空载体转染细胞 )及CHO细胞的DNA为模板 ,在PE5 70 0PCR仪上进行荧光定量PCR分析。每个样本共检测 30次 ,结果采用 q检验分析。【结果】PcDNA3 1(+) /BDNF/CHO细胞、PcDNA3 1(+) /CHO和CHO细胞株中BDNF的拷贝数分别为 95 16 4± 12 ,316 2 2± 10 ,316 2 2± 11。q检验结果显示前者与后两者之间有统计学意义 ,P <0 .0 5 ,而后两者之间无统计学意义 ,即 P >0 .0 5。PcDNA 3 1(+) /BDNF/CHO细胞株中BDNF的拷贝数是后二者的 3倍 ,而后两者的拷贝数相同。【结论】BDNF转染细胞株中外源BDNF基因是以 2个拷贝的比例整合到宿主细胞基因组中去的。
Objective To detect the copy numbers of brain derived neurotrophic factor (BDNF) gene in BDNF transfected PcDNA3 1(+)/BDNF/CHO cells with fluorescent quantitative polymerase chain reaction (FQ PCR). Methods BDNF DNA were amplified by GeneAmp 5700 Sequence Detection System with equal quantitative genomic DNA of PcDNA3 1(+)/BDNF/CHO, PcDNA3 1(+)/CHO and CHO cells as tamplates respectively. The process was repeated 30 times for every samples. The results were analyzed using q test. Results The copy numbers of BDNF of PcDNA3 1(+)/BDNF/CHO cells and PcDNA 3 1(+)/CHO and CHO cells were 95 164±12, 31 622±10, 31 622±11 respectively. The copy numbers of BDNF of PcDNA3 1(+)/BDNF/CHO cells were as three times as those of the PcDNA3 1(+)/CHO and CHO cells. The copy numbers of the two latters were the same. Conclusion The results clearly show that the PcDNA3 1(+)/BDNF/CHO cells harbor two BDNF DNA copies.
出处
《中山医科大学学报》
CSCD
北大核心
2001年第1期8-10,共3页
Academic Journal of Sun Yat-sen University of Medical Sciences
基金
国家技术创新项目! (国经贸技 1998-3 4 5 )
