Cloning and Expression Analysis on BoBURP2 Gene of Brassica oleracea var. italica
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摘要: BURP是植物特有蛋白，在生长发育和抗逆反应中起着重要作用。本研究以青花菜为材料，利用PCR克隆到1个BURP基因，命名为BoBURP2。测序结果表明，BoBURP2的基因组DNA全长为1 218 bp，具1个内含子，编码区全长为840 bp，编码279个氨基酸；序列比对结果表明，该基因与BURP家族成员RD22具有较高的同源性。系统发育分析结果表明，BoBURP2与甘蓝、芜菁和甘蓝型油菜的同源序列相似性最高，在发育树上聚为一组，与醉蝶花的相似性最低，亲缘关系最远。RT-PCR结果表明，BoBURP2的表达受NaCl和干旱胁迫的诱导，表达量分别在24 h和48 h时达到最大值，暗示该基因与这2种逆境响应相关。青花菜BoBURP2的克隆与表达分析，为后续基因功能的鉴定和抗逆分子育种奠定了基础。Abstract: A plant-specific protein, BURP plays an important role in the growth, development, and stress response of a plant. In this study, a BURP gene designated as BoBURP2 was isolated from broccoli, Brassica oleracea var. italica, using PCR. The gene sequencing showed its genome DNA was 1 218 bp with a single intron. The complete coding sequence was 840 bp in length, encoding 279 amino acids. It shared a high homology with RD22, which is also a member of the BURP family. Phylogenetic analysis on BoBURP2 showed its sequence to be highly similar to those of B. oleracea, B. rapa, and B. napus. They could possibly be grouped in a same clade. On the other hand, the low similarity it shared with the sequence of Tarenaya hassleriana suggested a remote relationship between them. The RT-PCR results indicated the expression of BoBRUP2 was induced by both NaCl and drought stresses, with the highest levels detected at 24 h and 48 h. The cloning and expression analysis on BoBURP2 provided the information for further studies on the identification of its genetic function as well as the molecular breeding for stress resistant B. oleracea var. italica.
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Key words:
- Brassica oleracea var. italica /
- BoBURP2 /
- RD22 /
- cloning /
- expression
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图 1 BoBURP2基因的编码区及推导的氨基酸序列
注：阴影部分为BURP结构域。
Figure 1. Coding sequence of BoBURP2 gene and its deduced amino acid sequence
图 2 BoBURP2与8种同源氨基酸的序列比对
Figure 2. Sequence alignments of BoBURP2 and 8 homologous amino acids
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