摘要
【目的】 克隆青蒿鲨烯合酶基因,为基因工程改造青蒿打下基础。【方法】 采用聚合酶链反应(PCR)扩增、逆转录聚合酶链反应(RT-PCR)扩增、扩增片段与T-载体连接和克隆片段的序列分析等方法。【结果】 通过PCR扩增和RT-PCR扩增,分别获得3 590 bp及1 257 bp片段,克隆后经PCR和酶切反应进行了鉴定。初步测序结果与GenBank上已登录的青蒿鲨烯合酶基因序列同源性为99％,氨基酸序列同源性为100％。【结论】 成功克隆了青蒿鲨烯合酶基因及其cDNA。
【Objective】 To clone the squalene synthase gene of Artemisia annua L. for improving the quality and production of Artemisia annua L. by genetic engineering. 【Methods】 PCR amplification, RT-PCR amplification, ligation of the target fragment with a T-vector and sequence analysis of the interested gene were performed. 【Results】 An expected 3590 bp fragment was amplified by PCR and an expected 1257 bp fragment was amplified by RT-PCR. The two cloned fragments were identified by PCR and restriction enzyme digestion respectively. The preliminary sequence data indicated that the results obtained were similar to that from GenBank, and the difference was only found in several base pairs. 【Conclusion】 The squalene synthase gene and cDNA of Artemisia annua L. were successfully cloned and sequenced.
出处
《广州中医药大学学报》
CAS
2004年第5期387-390,共4页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
国家自然科学基金(编号:30271591)
国家中医药管理局科研基金(编号:1020024)
广东省自然科学基金(编号:020799)
青蒿素科技基金(编号:qhs02a10)
关键词
青蒿
鲨烯合酶基因
DNA
重组
序列分析
DNA
ARTEMISIA ANNUA
SQUALENE SYNTHASE GENE
DNA, RECOMBINANT
SEQUENCE ANALYSIS
