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从紫花苜蓿中克隆和鉴定一个新的锌指蛋白编码基因

Molecular cloning and characterization of a novel gene encoding zinc finger protein from Medicago sativa L.

机构信息

Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, People's Republic of China.

出版信息

Mol Biol Rep. 2009 Nov;36(8):2315-21. doi: 10.1007/s11033-009-9450-5. Epub 2009 Jan 22.

Abstract

A suppression subtraction hybridization (SSH) cDNA library had been constructed to identify differentially expressed genes. Based on the sequence of an expressed sequence tag (EST) homologous to Pisum sativum zinc finger protein mRNA (Accession number: AF160911), the full-length cDNA of 1,676 nucleotides was cloned from alfalfa by rapid amplification of cDNA ends (RACE). It was designated as MsZFN, encoding a protein of 418 amino acids. The amino acid sequence compared by blast revealed high homology with zinc finger protein of other plants. Sequence comparison showed that there were five conserved typical zinc finger motifs, and one sugar transfer protein signature. The calculated molecular weight of the MsZFN protein was 45.8 k Da, and theoretical isoelectric point was 8.13. The MsZFN localized in nucleus. Under normal growth conditions, differential expression of MsZFN exhibited that the expression was the highest in leaf and the lowest in root. MsZFN was quickly and transiently induced by NaCl treatment and reached its maximum at 30 min.

摘要

为了鉴定差异表达基因,构建了一个抑制性消减杂交(SSH)cDNA 文库。根据豌豆锌指蛋白 mRNA(登录号:AF160911)的表达序列标签(EST)同源序列,通过快速扩增 cDNA 末端(RACE)从紫花苜蓿中克隆出全长为 1676 个核苷酸的 cDNA。它被命名为 MsZFN,编码一个 418 个氨基酸的蛋白质。通过blast 进行的氨基酸序列比较显示与其他植物的锌指蛋白具有高度同源性。序列比较表明存在五个保守的典型锌指模体和一个糖转移蛋白特征。MsZFN 蛋白的计算分子量为 45.8 kDa,理论等电点为 8.13。MsZFN 定位于细胞核内。在正常生长条件下,MsZFN 的差异表达表现为在叶中表达最高,在根中表达最低。MsZFN 被 NaCl 处理迅速且短暂诱导,在 30 分钟时达到最大值。

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