安徽农业科学
安徽農業科學
안휘농업과학
JOURNAL OF ANHUI AGRICULTURAL SCIENCES
2010年
10期
4996-4998,5073
,共4页
常晶%郭春华%尹永志%江明锋%喻麟%徐亚欧%郑玉才
常晶%郭春華%尹永誌%江明鋒%喻麟%徐亞歐%鄭玉纔
상정%곽춘화%윤영지%강명봉%유린%서아구%정옥재
青蒿%鲨烯合酶%序列分析
青蒿%鯊烯閤酶%序列分析
청호%사희합매%서렬분석
Artemisia apiacea%Squalene synthase%Sequence analysis
[目的]对青蒿鲨烯合酶进行cDNA克隆,并进行序列分析.[方法]根据GenBank上已发表的鲨烯合酶cDNA基因序列设计1对特异性引物,提取青蒿细胞总RNA,运用RT-PCR扩增出鲨烯合酶基因.将其与pMD19-T载体连接,并对克隆片段序列进行分析.[结果]SS基因全长1 257 bp,编码418个氨基酸;同源性分析表明,青蒿SS基因序列与人参、积雪草、金铁锁、大豆、辣椒、百脉根、远志、玉米、褐家鼠、小鼠以及人相应序列的同源性分别为77.21%、76.11%、75.66%、74.62%、73.83%、74.46%、73.03%、64.39%、52.22%、50.51%和50.12%;氨基酸同源性分别为79.43%、79.00%、75.84%、79.43%、77.27%、77.27%、77.03%、66.03%、40.65%、40.19%和40.09%.[结论]青蒿鲨烯合酶cDNA的成功克隆,为进一步研究SS基因结构、基因表达与调控提供了重要依据.
[目的]對青蒿鯊烯閤酶進行cDNA剋隆,併進行序列分析.[方法]根據GenBank上已髮錶的鯊烯閤酶cDNA基因序列設計1對特異性引物,提取青蒿細胞總RNA,運用RT-PCR擴增齣鯊烯閤酶基因.將其與pMD19-T載體連接,併對剋隆片段序列進行分析.[結果]SS基因全長1 257 bp,編碼418箇氨基痠;同源性分析錶明,青蒿SS基因序列與人參、積雪草、金鐵鎖、大豆、辣椒、百脈根、遠誌、玉米、褐傢鼠、小鼠以及人相應序列的同源性分彆為77.21%、76.11%、75.66%、74.62%、73.83%、74.46%、73.03%、64.39%、52.22%、50.51%和50.12%;氨基痠同源性分彆為79.43%、79.00%、75.84%、79.43%、77.27%、77.27%、77.03%、66.03%、40.65%、40.19%和40.09%.[結論]青蒿鯊烯閤酶cDNA的成功剋隆,為進一步研究SS基因結構、基因錶達與調控提供瞭重要依據.
[목적]대청호사희합매진행cDNA극륭,병진행서렬분석.[방법]근거GenBank상이발표적사희합매cDNA기인서렬설계1대특이성인물,제취청호세포총RNA,운용RT-PCR확증출사희합매기인.장기여pMD19-T재체련접,병대극륭편단서렬진행분석.[결과]SS기인전장1 257 bp,편마418개안기산;동원성분석표명,청호SS기인서렬여인삼、적설초、금철쇄、대두、랄초、백맥근、원지、옥미、갈가서、소서이급인상응서렬적동원성분별위77.21%、76.11%、75.66%、74.62%、73.83%、74.46%、73.03%、64.39%、52.22%、50.51%화50.12%;안기산동원성분별위79.43%、79.00%、75.84%、79.43%、77.27%、77.27%、77.03%、66.03%、40.65%、40.19%화40.09%.[결론]청호사희합매cDNA적성공극륭,위진일보연구SS기인결구、기인표체여조공제공료중요의거.
[Objective] cDNA from squalene synthase was cloned and sequenced. [Method] A pair of specific primers were designed according to the cDNA gene sequence of squalene synthase published in GenBank. Total RNA was extracted from the cell of Artemisia apiacea. The genes of squalene synthase were amplified by using RT-PCR. It was connected with pMD19-T vector and the cloned fragment sequences were analyzed. [Result] SS gene with the whole length of 1257 bp was amplified and the fragment encoded 418 amino acids. The homology of SS gene between Artemisa apiacea and Panax ginseng, Centella asiatica, Psammosilene tunicoides, Glycine max, Capsicum annuum, Lotus japonicus, Polygala tenuifolia, Zea mays, Rattus norvegicus, Mus musculus and human were 77.21%, 76.11%, 75.66%, 77.27%, 77.03%, 66.03%, 40.65%, 40.19% and 40.09%, respectively. [Conclusion] cDNA of squalene synthase from Artemisia apiacea was successfully cloned, which provided important base for further study on the structure, gene expression and regulation of SS gene.