鹰嘴豆种子中凝集素基因的分离与鉴定。

Insaf A Qureshi, Prasanta K Dash, Prem S Srivastava, Kirpa R Koundal
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引用次数: 9

摘要

以西芹(Cicer arietinum)未成熟种子的poly (A(+)) RNA为载体,在lambda TriplEx2载体上构建cDNA文库。通过文库筛选和RACE-PCR技术从鹰嘴豆种子中分离到凝集素基因。豌豆种子凝集素(CpGL)全长972 bp,包含一个807 bp的开放阅读框,编码268个氨基酸的蛋白。分析表明,CpSL基因与其他豆科凝集素基因具有较强的同源性。系统发育分析表明,CpSL存在两个主要聚类,明确属于甘露糖特异性凝集素家族。RT-PCR结果显示,CAA基因在花、叶、根、茎等多种植物组织中均有组成性表达。在发育中的鹰嘴豆凝集素mRNA水平检测中,10个DAF种子的凝集素mRNA水平较高,在整个种子发育过程中呈下降趋势。
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Isolation and characterization of a lectin gene from seeds of chickpea (Cicer arietinum L.).

A cDNA library was constructed in lambda TriplEx2 vector using poly (A(+)) RNA from immature seeds of Cicer arietinum. The lectin gene was isolated from seeds of chickpea through library screening and RACE-PCR. The full-length cDNA of Chichpea seed lectin(CpGL)is 972 bp and contains a 807 bp open reading frame encoding a 268 amino acid protein. Analysis shows that CpSL gene has strong homology with other legume lectin genes. Phylogenetic analysis showed the existence of two main clusters and clearly indicated that CpSL belonged to mannose-specific family of lectins. RT-PCR revealed that CAA gene expressed constitutively in various plant tissues including flower, leaf, root and stem. When chickpea lectin mRNA level was checked in developing seeds, it was higher in 10 DAF seeds and decreased throughout seed development.

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