农业科学与技术（英文版）
農業科學與技術（英文版）
농업과학여기술（영문판）
AGRICULTURAL SCIENCE & TECHNOLOGY
2010年
2期
80-83
,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,并进行其序列分析.[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,并通过DNAman、NCBI Blast、ExPASy ProtParam等工具,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%、77.03%、66.03%、40.65%、40.19%和40.09%.由SS基因翻译出来的蛋白质分子量为47.839 4 KDa,理论等电点为8.57,无信号肽.疏水性分析表明,在400～417区域疏水性较强.跨膜区分析表明有3处跨膜区域,分别为170～186位的17个氨基酸、282～305位的24个氨基酸、387～407位的21个氨基酸.通过NCBI的BLAST程序分析指出,该基因编码的氨基酸序列含有Trans_IPPS_HH的保守区域,也具有与Mg2+结合有关的活性中心--富含天门冬氨酸的区域(DXXDD).2级结构以α-螺旋和无规则卷曲为主.[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,併進行其序列分析.[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,併通過DNAman、NCBI Blast、ExPASy ProtParam等工具,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%、77.03%、66.03%、40.65%、40.19%和40.09%.由SS基因翻譯齣來的蛋白質分子量為47.839 4 KDa,理論等電點為8.57,無信號肽.疏水性分析錶明,在400～417區域疏水性較彊.跨膜區分析錶明有3處跨膜區域,分彆為170～186位的17箇氨基痠、282～305位的24箇氨基痠、387～407位的21箇氨基痠.通過NCBI的BLAST程序分析指齣,該基因編碼的氨基痠序列含有Trans_IPPS_HH的保守區域,也具有與Mg2+結閤有關的活性中心--富含天門鼕氨痠的區域(DXXDD).2級結構以α-螺鏇和無規則捲麯為主.[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,병진행기서렬분석.[방법]근거GenBank상이발표적사희합매cDNA기인서렬설계1대특이성인물,제취청호세포총RNA,운용RT-PCR확증출사희합매기인.장기여pMD19-T재체련접,병통과DNAman、NCBI Blast、ExPASy ProtParam등공구,대극륭편단서렬진행분석.[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%、77.03%、66.03%、40.65%、40.19%화40.09%.유SS기인번역출래적단백질분자량위47.839 4 KDa,이론등전점위8.57,무신호태.소수성분석표명,재400～417구역소수성교강.과막구분석표명유3처과막구역,분별위170～186위적17개안기산、282～305위적24개안기산、387～407위적21개안기산.통과NCBI적BLAST정서분석지출,해기인편마적안기산서렬함유Trans_IPPS_HH적보수구역,야구유여Mg2+결합유관적활성중심--부함천문동안산적구역(DXXDD).2급결구이α-라선화무규칙권곡위주.[결론]청호사희합매cDNA적성공극륭,위진일보연구SS기인결구、기인표체여조공제공료중요의거.
[Objective] cDNA from squalene synthase was cloned and sequenced. [Method] A pair of specific primers was 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 1 257 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%, 74.62%, 73.83%, 74.46%, 73.03%, 64.39%, 52.22%, 50.51% and 50.12%, respectively. The homology of the deduced amino acid were 79.43%, 79.00%, 75.84%, 79.43%, 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.
