食品科学

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甜杏仁和苦杏仁野黑樱苷水解酶基因的克隆

白羽嘉1,2,王 敏2,陶永霞2,徐 乐3,冯作山1,2,*   

  1. 1.新疆农业大学林学与园艺学院,新疆 乌鲁木齐 830052;2.新疆农业大学食品科学与药学学院,新疆 乌鲁木齐 830052;
    3.新疆农业科学院轮台果树资源圃,新疆 轮台 841600
  • 发布日期:2014-12-15

Cloning of Prunasin Hydrolase Gene from Sweet and Bitter Apricot Kernels

BAI Yu-jia1,2, WANG Min2, TAO Yong-xia2, XU Le3, FENG Zuo-shan1,2,*   

  1. 1. College of Forestry and Horticulture, Xinjiang Agricultural University, Ürümqi 830052, China;
    2. College of Food Science and Pharmacy, Xinjiang Agricultural University, Ürümqi 830052, China;
    3. Luntai Plant Germplasm Resources Garden, Xinjiang Academy of Agricultural Sciences, Luntai 841600, China
  • Published:2014-12-15

摘要:

以轮台小白杏(甜杏仁品种)和甲麦黄杏(苦杏仁品种)为实验材料,采用同源基因克隆方法克隆野黑樱苷水解酶(prunasin hydrolase,PH)基因。轮台小白杏PH基因命名为PaLTPh(GenBank登录号:KF888615),序列长度3 686 bp;甲麦黄杏PH基因命名为PaMJPh(KF888616),序列长度3 690 bp。PaLTPh和PaMJPh与扁桃Ph691基因相似性为94%。根据PaLTPh和PaMJPh保守区序列设计引物,克隆编码区(coding sequence,CDS)序列。PaLTPh CDS(KF888617)和PaMJPh CDS(KF888618)序列长度均为1 635 bp,与扁桃Ph691全CDS区相似性为97%。PaLTPh CDS和PaMJPh CDS序列均为全长的开放阅读框,编码蛋白质长度为544 个氨基酸。PaLTPH蛋白和PaMJPH蛋白均包含长度为26 个氨基酸的信号肽和糖基水解酶家族Ⅰ结构域,可催化野黑樱苷水解为扁桃腈和葡萄糖。杏PH基因与扁桃Ph691具有高度的同源性。

关键词: 轮台小白杏, 甲麦黄杏, 野黑樱苷, 野黑樱苷水解酶, 基因克隆

Abstract:

Prunasin hydrolase (PH) gene was cloned from sweet and bitter apricot (Prunus armeniaca L.) kernels from
the cultivars Luntai Xiaobaixing and Jiamaihuangxing, respectively. The PH gene from Luntai Xiaobaixing was named as
PaLTPh (GenBank accession number: KF888615) with a sequence length of 3 686 bp, and that from Jiamaihuangxing was
named as PaMJPh (GenBank accession number: KF888616) with a sequence length of 3 690 bp. Analysis of the nucleotide
sequences indicated that the homology between the PH gene and the PH691 gene of Prunus dulcis reached up to 94%.
Moreover, the coding sequence (CDS) region was cloned according to the conserved regions of PaLTPh and PaMJPh. The
sequence length of both PaLTPh CDS (KF888617) and PaMJPh CDS (KF888618) was 1 635 bp. Both these CDS regions
were full open reading frame (ORF) sequences, encoding a protein consisting of 544 amino acid (aa) residues. Both proteins
contained an amino-terminal signal peptide with 26 AA residues and the domain of glycosyl hydrolase family Ⅰ for PaLTPH
and PaMJPH, which can catalyze the hydrolysis of prunasin to mandelonitrile and glycosides. This study has demonstrated
that the PH gene from apricot has high homology with the Ph691 of Prunus dulcis.

Key words: Prunus armeniaca L. cultivar Luntai Xiaobaixing, Prunus armeniaca L. cultivar Jiamaihuangxing, prunasin, prunasin hydrolase, gene cloning

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