摘要
根据已知的其他物种PNAE酶cDNA序列设计引物,采用RT-PCR技术从云南萝芙木叶片中扩增获得pnae基因部分cDNA序列即PNAE酶基因中间大片段,再用RACE技术获得其两端序列。序列拼接得到完整的1 004 bp的PNAE酶基因,根据获得的序列,分析得到795 bp的开放阅读框,编码264个氨基酸。序列分析显示,云南萝芙木中PNAE酶氨基酸序列与蛇根木中的该酶氨基酸序列同源性高达90%,但和其他植物物种中的PNAE酶氨基酸序列,以及其他物种间的PNAE酶氨基酸序列同源性都不高,在40%-60%之间,表明不同物种中PNAE酶氨基酸序列不具有全序列的高度同源性。进一步序列分析发现,在各植物的PNAE酶氨基酸序列中都存在两个高度保守的氨基酸区域,表明不同物种中PNAE酶存在共同的高度保守区段。
According to some identified cDNA sequence of the polyneuridine aldehyde esterase （ PNAE ） gene in plant species on the NCBI, pairs of primers were designed, and the partial intermediate sequence of the PNAE cDNA was obtained using RT-PCR technique in Rauvolfia yunnanensis＇ fresh leaves, and then by the technique called rapid-amplification of CDNA ends （ RACE ）, the sequence in both two flanks of the known sequence was acquired. The full length sequence of the cDNA of PNAE in Rauvolfia yunnanensis was I 004 bp and that included an open reading frame （ ORF ） which encoded 264 amino acids. Sequence analysis showed that for PNAE gene the homologies of amino acid sequence among each species was a little low ranging from 40% to 60%, demonstrating PNAE gene wasn＇ t a gene that its complete sequence is relatively conservative. The further analysis of all the known PNAE in plant species found that there are two conservative domains in amino acid sequence, revealing that PNAE gene has two highly conserved domains among different plant species.
出处
《生物技术通报》
CAS
CSCD
北大核心
2013年第4期75-80,共6页
Biotechnology Bulletin
基金
云南2007年度产业化重大关键技术开发项目[云发改高科(2007)1718号]
中南林业科技大学校级学科重点建设项目(066)
