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烟草中一条新的S-腺苷甲硫氨酸合成酶基因的克隆及表达分析

Cloning and Characterization of a Novel S-Adenosyl-L-Methionine Synthase Gene in Tobacco

  • 摘要: 利用差异显示技术从烟草中分离到一个受乙烯利诱导表达的cDNA片段,结合RACE扩增技术获得该基因的全长序列。该全长cDNA共含1350个碱基,编码区有1170bp,通过GenBank同源性比较发现它与一条已知的烟草S-腺苷甲硫氨酸合成酶(SAMS1,EC2.5.1.6)的基因编码区存在90%的同源性,编码的氨基酸顺序96%同源,而在cDNA的5’及3’非编码区同源性很低,且有一些短片段的缺失。设计合适的引物分别对这两个SAMS基因进行RT-PCR分析表明,SAMS1基因受高温诱导表达增强,但不受甲基紫晶诱导的氧化胁迫及盐胁迫影响,而新分离到的这个基因(SAMS2)的表达同时受高温、甲基紫晶及高盐3种胁迫的诱导。

     

    Abstract: Differential display method was used to identify novel genes regulated by the ethylene generator,ethephon in tobacco plants.A full length cDNA of S-Adenosyl-L-Methionine synthase (SAMS2) which contains 1350 bp was isolated by this way combined with RACE technology.Sequence analysis revealed that this gene shared 90% similarity in gene sequence and 96% similarity in protein sequence with a known SAMS (SAMS1) gene though they have low homologous in the 5’ and 3’ uncoden region.RT-PCR analysis indicated that the expression of SAMS1 was induced by the treatment of high temperature but has no response to oxidative stress induced by methyl viologen (MV) and NaCl stress,while SAMS2 was up-regulated by all the three stress treatment.

     

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