All Title Author
Keywords Abstract

-  2017 

枸杞苯丙氨酸解氨酶基因的克隆与表达分析
Molecular cloning and expression patterns of LcPAL from Lycium chinense

Keywords: 枸杞 苯丙氨酸解氨酶(PAL) 基因克隆 表达分析
Lycium chinense phenylalanine ammonia-lyase(PAL) gene cloning expression analysis

Full-Text   Cite this paper   Add to My Lib

Abstract:

为研究枸杞苯丙氨酸解氨酶基因LcPAL在干旱和盐逆境中的作用,采用RT-PCR和RACE方法克隆了LcPAL基因(Genbank No.KX781247)。该基因cDNA长度为2 544 bp,含有2 163 bp的完整开放阅读框,编码720个氨基酸。蛋白序列分析表明,其包含典型的PAL酶活性中心序列(GTITASGDLVPLSYIA),与其他植物的PAL蛋白有很高的同源性。系统进化树表明,枸杞LcPAL与茄科植物的PAL蛋白聚为一类,说明两者的亲缘关系较近。利用实时荧光定量PCR方法检测发现,LcPAL基因在枸杞的叶中表达量最高,果实中表达量最少。其在聚乙二醇(PEG)、氯化钠(NaCl)胁迫下诱导表达,但表达模式不同。研究结果推测从枸杞中克隆获得苯丙氨酸解氨酶(LcPAL),是典型的PAL家族成员,在枸杞各组织发育过程中具有重要功能,且在枸杞抗干旱和抗盐逆境过程也起一定作用。
To investigate the role of LcPAL from Lycium chinense in drought and salt stress,a phenylalanine ammonia-lyase(PAL)gene was cloned from Lycium chinense by the methods of reverse-transcription PCR and RACE.The full-length cDNA of L.chinense PAL (designated as LcPAL)was 2 544 bp(Genbank No.KX781247)containing a complete open reading frame(ORF)of 2 163 bp,which encoded 720 amino acid residues.Sequence analysis showed that it had typical PAL enzyme active site sequence(GTITASGDLVPLSYIA).Homology analysis indicated that the deduced LcPAL protein was highly homology to other PAL proteins from different species.Phylogenetic tree analysis revealed that LcPAL had closer relationship with PALs from Solanaceae plants than from other plants.The results from Real-Time PCR indicated that the epression of LcPAL gene was the strongest in the leaves of L.chinense,and the least in fruits.Furthermore,LcPAL transcription level was induced under the treatment of PEG and NaCl stresses,but the expression patterns were different.As a result,the gene LcPAL which was cloned and characterized from L.chinense is a member of PAL family typically.The result of study indicated that LcPAL played a role in the development of L.chinense plant.LcPAL also played a role in resistance to drought and salt stress processes.

Full-Text

comments powered by Disqus