【摘要】 将ｂｄｎｆ基因克隆入逆转录病毒载体ｐＬＮＣＸ，构建得到ｐＬＮＣ／ＢＤＮＦ，经ＰＡ３１７细胞包装后，感染大鼠成肌细胞Ｌ６ＴＧ，Ｇ４１８筛选２周后，得到稳定表达ｂｄｎｆ基因的细胞克隆Ｌ６ＴＧ／ＢＤＮＦ。ＤＮＡ印迹结果证实ｂｄｎｆ基因已经整合入Ｌ６ＴＧ染色体中，ＲＮＡ印迹和斑点印迹结果分别从ｍＲＮＡ水平和蛋白水平证明了ｂｄｎｆ基因的表达，且Ｌ６ＴＧ／ＢＤＮＦ培养上清中ＢＤＮＦ的含量约为２５ｎｇ（１０６细胞数每ｍｌ每２４ｈ）。更多还原

【Abstract】 Brain derived neurotrophic factor (BDNF) gene was cloned into retrovirous vector plasmid pLNCX to form the pLNC/BDNF. After packaging in PA317 cell line, the infectious particles were used to infect rat myoblast cell line L 6TG. After selection with G418, the L 6TG/BDNF cells was collected. Results from Southern blot show that bdnf gene has successfully integrated into the L 6TG genomic DNA. The expression of bdnf gene has been proved by Northern blot and Dot blot. The content of BDNF in the culture medium of L 6TG/BDNF was about 25 ng·10 -6 cells per 24 h per ml.更多还原