摘要
为了转录出反义ｂｃｒ／ａｂｌＲＮＡ，我们用逆转录。聚合酶链反应技术将ｂｃｒ／ａｂｌ嵌合转录体接合区扩增，并反向插到ｐＧＥＭ－４Ｚ质粒ＳＰ６ＲＮＡ聚合酶转录起始点下游作为体外转录的模板。限制酶切分析和ＤＮＡ序列分析结果都证实插入片段序列与Ｋ５６２ｍＲＮＡ接合区完全一致。利用体外转录技术，我们成功地转录出跨ｂｃｒ／ａｂｌ接合区反义ＲＮＡ。体外杂交实验的结果表明，该反义ＲＮＡ具有杂交捕获ｂｃｒ／ａｂｌｍＲＮＡ接合区的功能，可应用于选择性抑制含ｂｃｒ／ａｂｌ基因的白血病细胞。
o transcribe the antisense RNA of bcr/abl gene, weamplified the junction region of the bcr/abl chimerictranscripts by RT/PCR technique and inserted it intoPGEM-4Z plasmid down stream the SP6 RNA poly-merase transcription initiation site in inverted directionas templet. The results of restriction analysis and sequencing analysis confirmed that the sequence of the0. 3kb inserted fragment were exactly the same sa K562mRNA junction region. The antisense RNA of bcr/ablfusion gene was successfully transcribed following the invtiro transcription protocol. The results of in vitro hy-bridization experiments demonstrated that this antisenseRNA had tho ability to arrest bcr/abl mRNA at the junc-tion region. Is can be used in furrher studies on selectivesuppression of ph￣+ leukemic cells.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
1994年第9期469-471,共3页
Chinese Journal of Hematology
基金
天津市21世纪青年科学基金
关键词
反义RNA
基因
杂交
白血病
转录
Antisense RNA bcr/abl gene Invitro transcription Hybridization
