摘要
Δ′-吡咯啉-5-羧酸合成酶(P5CS)是植物渗透胁迫下脯氨酸合成谷氨酸的关键酶。利用RACE结合RT-PCR技术克隆获得大蒜芥(Sisybrium altissimum)SaP5CS1基因全长cDNA序列,该基因全长1 907bp,其中开放阅读框为1 719bp,编码573个氨基酸,预测编码蛋白质的等电点为4.95,分子量为156.279kDa。同源分析显示,SaP5CS1与甘蓝型油菜(Brassica napus)和拟南芥(Arabidopsis thaliana)的蛋白序列相似性分别为98%和96%。实时荧光定量PCR分析表明,在干旱逆境胁迫条件下,大蒜芥SaP5CS1基因在根和叶的相对表达量受胁迫的诱导均上调。SaP5CS1基因的克隆及序列分析将为其功能分析和分子育种奠定基础。
Delta′-pyrroline-5-carboxylatesynthetase(P5CS)is the key enzyme for glutamic acid biosynthesis from proline under plant osmotic stress.In the present study,the full-length complementary DNA(cDNA)sequence of one P5CSgene(designated SaP5CS1)in Sisybrium altissimum was characterized by RACE and real-time PCR(RT-PCR).The complete cDNA sequence of SaP5CS1 comprised 1 907 bp with an open reading frame(ORF)of 1 719 bp,and encoded a P5 CSprecursor peptide of 573 amino acids with apredicted molecular weight of 156.279 kDa and isoelectric point of 4.95.In the sequence alignments of P5 CSgenes,the amino acid sequences of SaP5CS1 exhibited 98%and 96%sequence identities with Brassica napus P5CS1 and Arabidopsis thaliana P5CS1,respectively.Real-time fluorescent quantitative PCR(qRT-PCR)analysis showed that SaP5CS1 gene was highly up-regulated in both leaves and roots at different drought stress treatment time points.The cloning and sequence analysis of SaP5CS1 gene might lay a foundation for the further function analysis and molecular breeding.
出处
《草业科学》
CAS
CSCD
北大核心
2015年第2期210-216,共7页
Pratacultural Science
基金
国家转基因生物新品种重大专项"转基因耐旱
耐盐新品种培育"(2013ZX08005-004)
关键词
大蒜芥
P5CS
相对表达量
序列分析
Sisybrium altissimum
P5CS
relative expression
sequence analysis
