摘要
用逆转录聚合酶链反应技术从Ｋ５６２细胞系中扩增出ｂｃｒ／ａｂ１融合区２５３ｂｐＤＮＡ片段，经反复连接与克隆，得到含同方向串联的２、３和４个拷贝的该融合基因片段的克隆载体ｐＵＣｓ。这些片段分别反向连接到逆转录病毒表达载体ｐＤＯＲｎｅｏ。ＤＮＡ序列分析、限制性内切酶分析和菌落原位杂交证实：不同拷贝数的ｂｃｒ／ａｂ１融合区基因片段已反向插入ｐＤＯＲｎｅｏ。利用脂质体介导，将重组质粒导入Ｋ５６２细胞系后。
The 253 bp bcr/ab1 fusion gene was obtained from K562 cell line by reverse transcriptionpolymerase chain reaction (RTPCR). Through repetitive ligations and clonings, the cloning vectors pUCs containing two, three and four copies of the fragments with the same orientation were obtained. These fragments were further inserted into retroviral vector pDORneo in antisense direction. DNA sequencing, electrophoresis and hybridization showed that bcr/ab1 fragments with different copies had been inserted into pDORneo as designed. When the K562 cell line was transfected with the recombinant plasmids by lipofectin reagent, inhibition of proliferation of the transfected cells was observed.
出处
《军事医学科学院院刊》
CSCD
北大核心
1996年第2期107-109,共3页
Bulletin of the Academy of Military Medical Sciences
