Detection of the Copy Numbers of Exogenous Gene of Transfected Cells by Fluorescent Quantitative Polymerase Chain Reaction
- VernacularTitle:用荧光定量PCR方法检测转染细胞中外源基因的拷贝数
- Author:
Ying LI
;
Yunshao HE
;
Gang CHENG
;
Hu LI
- Publication Type:Journal Article
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2001;22(1):14-18
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To detect the copy numbers of brain-deriv ed neurotrophic factor (BDNF) gene in BDNF transfected PcDNA3.1(+)/BDNF/CHO cel ls with fluorescent quantitative polymerase chain reaction (FQ-PCR). 【Methods 】 BDNF DNA were amplified by GeneAmp 5700 Sequence Detection System with eq ual 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 sam ples. The results were analyzed using q test. 【Results】 The copy numbers o f BDNF of PcDNA3.1(+)/BDNF/CHO cells and PcDNA 3.1(+)/CHO and CHO cells were 9 5 164±12, 31 622±10, 31 622±11 respectively. The copy numb ers of BDNF of PcDNA3.1(+)/BDNF/CHO cells were as three times as those of the P cDNA3.1(+)/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 h arbor two BDNF DNA copies.
