| 扁桃AcSFB基因的克隆与原核表达分析 |
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| 引用本文： | 郭长奎,罗淑萍,李疆,高启明.扁桃AcSFB基因的克隆与原核表达分析[J].园艺学报,2009,36(8):1215-1220. |
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| 作者姓名： | 郭长奎 罗淑萍 李疆 高启明 |
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| 作者单位： | 郭长奎;罗淑萍;李疆;高启明 |
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| 基金项目： | 国家自然科学基金，国家科技支撑计划 |
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| 摘 要： | 利用RT-PCR和RACE技术, 从新疆栽培扁桃‘鹰嘴’花药中获得了一个全长的SFB ( S-hap-lotype-specific F-box gene) 基因(GenBank登录号为FJ362525) , 命名为AcSFBd。该基因全长1 125 bp, 编码374个氨基酸, 在N末端含有一个大约50个氨基酸组成的F2box基序, 含有两个高变区(Hva和Hvb)和两个可变区(V1和V2) 。生物信息学的方法成功预测了AcSFBd蛋白分子质量为43 kD, 为亲水性、非分泌型蛋白, 在基质内起连接酶(EC6. - . - . -) 的作用。成功构建了原核表达载体, 并转化大肠杆菌BL21 (DE3) , SDS2PAGE检测结果显示表达了一个约60 kD的蛋白。
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| 关 键 词： | 扁桃 基因 克隆 SFB 基因 原核表达 生物信息学 |
| 收稿时间： | 2009-2-24 |
| 修稿时间： | 2009-7-2 |
Identification and Prokaryotic Expression of Almond AcSFB Gene |
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| GUO Chang-kui,LUO Shu-ping,LI Jiang,GAO Qi-ming.Identification and Prokaryotic Expression of Almond AcSFB Gene[J].Acta Horticulturae Sinica,2009,36(8):1215-1220. |
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| Authors: | GUO Chang-kui LUO Shu-ping LI Jiang GAO Qi-ming |
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| Institution: | (College of Agronomy, Xinjiang Agricultural University, Urumqi 830052, China; College of Forestry and Horticulture, Xinjiang Agricultural University, Urumqi 830052, China) |
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| Abstract: | Style S-determinants have been determined to be S-RNase and the SFB ( S-haplotype-specific F-box gene) alleles are considered to be the first candidates of the pollen S-determinants. A total length SFB gene (GenBank accession No1FJ362525) named AcSFBd was cloned with RT-PCR and RACE technolgy from the anther of almond Yingzui p lanted in Xinjiang. AcSFBd was 1 125 bp in length encoding a protein of 374 amino acids. This gene encoded proteins with the F-box motif in the N amino terminus, and with four ( hyper) variable regions, i e. , V1, V2, Hva and Hvb which were similarwith S-RNase genes, considered to be recognition site between SFB and S-RNase. With the bioinformatics analysis, the AcSFBd protein was 43 kD,was instable, hydrophilicity and non-secretory protein and had ligase enzyme activity. After transformed to Escherichia coli BL21 and induced by isopropyl-β-D-thiogalactopyranoside ( IPTG) , recombinant protein about60 kD was expressed in pET-AcSFB system and separated by SDS-PAGE electrophoresis |
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| Keywords: | almond gene clone SFB gene p rokaryotic exp ression bioinformatics |
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