浙江农业学报 ›› 2023, Vol. 35 ›› Issue (4): 831-840.DOI: 10.3969/j.issn.1004-1524.2023.04.10
收稿日期:
2021-12-13
出版日期:
2023-04-25
发布日期:
2023-05-05
通讯作者:
*闫会转,E-mail: hzhyan1118@163.com
作者简介:
燕存尧(1997—),男,新疆昌吉人,硕士研究生,主要从事蔬菜栽培与生理研究。E-mail: 1033146390@qq.com
基金资助:
YAN Cunyao(), JIA Kai, YAN Huizhuan(), GAO Jie
Received:
2021-12-13
Online:
2023-04-25
Published:
2023-05-05
摘要:
脂氧合酶(lipoxygenase,EC 1.13.11.12;简称LOX),是一类由非血红素铁组成的脂肪酸双加氧酶,在植物的生长发育和环境胁迫响应中发挥着重要作用,本研究以4片真叶期芜菁幼苗为材料,利用同源克隆技术从芜菁中克隆BrrLOX7基因。利用生物信息学方法分析BrrLOX7基因序列。使用荧光定量PCR技术分析非生物胁迫下BrrLOX7基因表达模式。结果显示,克隆的BrrLOX7基因CDS长度为2 715 bp,编码904个氨基酸,编码的蛋白质分子量为103.10 ku,无跨膜结构和信号肽。亚细胞定位预测显示,该基因编码的蛋白位于叶绿体中,该蛋白与白菜和萝卜的序列同源性和遗传距离最近。不同胁迫下的表达模式显示,BrrLOX7基因参与对4种胁迫(盐胁迫、冷胁迫、热胁迫、干旱胁迫)的响应。研究结果将为BrrLOX7基因的功能研究奠定基础。
中图分类号:
燕存尧, 贾凯, 闫会转, 高杰. 芜菁BrrLOX7基因克隆、表达及生物信息学分析[J]. 浙江农业学报, 2023, 35(4): 831-840.
YAN Cunyao, JIA Kai, YAN Huizhuan, GAO Jie. Cloning, expression and bioinformatics analysis of BrrLOX7 gene in turnip[J]. Acta Agriculturae Zhejiangensis, 2023, 35(4): 831-840.
图1 BrrLOX7反转录PCR M,DL5000 marker;1和2表示BrrLOX7基因的PCR扩增产物。
Fig.1 RT-PCR of BrrLOX7 M, DL5000 marker; 1 and 2 indicate PCR amplification products of the BrrLOX7 gene.
氨基酸的种类 Types of amino acid | 氨基酸的数目 Number of amino acids | 占氨基酸总量的百分比 Percent of total amino acids/% | 氨基酸的种类 Types of amino acid | 氨基酸的数目 Number of amino acids | 占氨基酸总量的百分比 Percent of total amino acids/% |
---|---|---|---|---|---|
苯丙氨酸Phe | 34 | 3.76 | 脯氨酸Pro | 62 | 6.86 |
亮氨酸Leu | 89 | 9.85 | 苏氨酸Thr | 45 | 4.98 |
异亮氨酸Ile | 47 | 5.20 | 丙氨酸Ala | 51 | 5.64 |
甲硫氨酸Met | 16 | 1.77 | 酪氨酸Tyr | 34 | 3.76 |
缬氨酸Val | 53 | 5.86 | 组氨酸His | 22 | 2.43 |
半胱氨酸Cys | 12 | 1.33 | 谷氨酰胺Gln | 25 | 2.77 |
色氨酸Trp | 21 | 2.32 | 天冬氨酸Asn | 30 | 3.32 |
精氨酸Arg | 49 | 5.42 | 赖氨酸Lys | 63 | 6.97 |
丝氨酸Ser | 60 | 6.64 | 天冬氨酸Asp | 61 | 6.75 |
甘氨酸Gly | 55 | 6.08 | 谷氨酸Glu | 75 | 8.30 |
表1 BrrLOX7基因编码蛋白的氨基酸组成
Table 1 Amino acid composition of the protein encoded by BrrLOX7 gene
氨基酸的种类 Types of amino acid | 氨基酸的数目 Number of amino acids | 占氨基酸总量的百分比 Percent of total amino acids/% | 氨基酸的种类 Types of amino acid | 氨基酸的数目 Number of amino acids | 占氨基酸总量的百分比 Percent of total amino acids/% |
---|---|---|---|---|---|
苯丙氨酸Phe | 34 | 3.76 | 脯氨酸Pro | 62 | 6.86 |
亮氨酸Leu | 89 | 9.85 | 苏氨酸Thr | 45 | 4.98 |
异亮氨酸Ile | 47 | 5.20 | 丙氨酸Ala | 51 | 5.64 |
甲硫氨酸Met | 16 | 1.77 | 酪氨酸Tyr | 34 | 3.76 |
缬氨酸Val | 53 | 5.86 | 组氨酸His | 22 | 2.43 |
半胱氨酸Cys | 12 | 1.33 | 谷氨酰胺Gln | 25 | 2.77 |
色氨酸Trp | 21 | 2.32 | 天冬氨酸Asn | 30 | 3.32 |
精氨酸Arg | 49 | 5.42 | 赖氨酸Lys | 63 | 6.97 |
丝氨酸Ser | 60 | 6.64 | 天冬氨酸Asp | 61 | 6.75 |
甘氨酸Gly | 55 | 6.08 | 谷氨酸Glu | 75 | 8.30 |
图4 BrrLOX7蛋白二级结构预测 蓝色代表 α-螺旋; 红色代表延伸链; 绿色代表 β-折叠; 紫色代表无规则卷曲。
Fig.4 Secondary structure of BrrLOX7 protein Blue for α-helix; red for extended chain; green for β-turn; purple for irregularly curled.
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