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为探索百合的耐盐,耐寒和耐旱的抗性机理,本试验选取了抗寒能力极强的岷江百合(Lilium regale)为材料,利用RT-PCR技术克隆得到了岷江百合的Li GPAT基因的ORF区序列,全长1 212 bp,编码403个氨基酸,分子量约为45.4 k D,Gen Bank登录号为JX524740。通过同源序列比对发现,Li GPAT基因编码的氨基酸与单子叶植物油棕GPAT基因编码的氨基酸的一致性最高,达到67.17%。利用在线软件SMART进行分析,表明Li GPAT氨基酸在169~315序列存在一个Pls C结构域,这就说明Li GPAT蛋白属于酰基转移酶家族成员,且所有的活性位点都位于这个区域,具有酰基转移酶的活性。通过在线软件TMHMM Server 2.0预测分析揭示Li GPAT氨基酸序列不存在跨膜区域,可能是一个可溶性蛋白。利用在线软件Plant-m PLoc预测表明,Li GPAT蛋白质的亚细胞定位为叶绿体。
In order to explore the mechanism of resistance to salt, cold and drought of lily, we selected the Lilium regale with strong cold-resistance ability as material and cloned the Li GPAT gene of Minjiang lily by RT- The ORF region was 1 212 bp in length and encoded 403 amino acids with a molecular weight of 45.4 kD and Gen Bank accession number JX524740. By homologous sequence alignment, it was found that the amino acids encoded by Li GPAT gene and the GPAT gene of monocot palm oil had the highest identity (67.17%). Analysis using on-line software SMART showed that there is a Pls C domain in the 169-115 sequence of Li GPAT amino acids, indicating that the Li GPAT protein is a member of the acyltransferase family and that all active sites are located in this region with acyl transfer Enzyme activity. Prediction analysis of online software TMHMM Server 2.0 reveals that the amino acid sequence of Li GPAT is not a transmembrane domain and may be a soluble protein. Plant-m PLoc predictions using online software showed that the subcellular localization of the Li GPAT protein is chloroplast.