|本期目录/Table of Contents|

[1]丁国林,胡宗利,陈国平,等.类球红细菌LHⅡβ-亚基/α-亚基融合蛋白吸收光谱分析[J].应用与环境生物学报,2013,19(03):395-398.[doi:10.3724/SP.J.1145.2013.00395]
 DING Guolin,HU Zongli,CHEN Guoping,et al.Absorption Spectral Analysis of Fused β-subunit and α-subunit Protein of Light-harvesting Ⅱ Complexes of Rb. sphaeroides[J].Chinese Journal of Applied & Environmental Biology,2013,19(03):395-398.[doi:10.3724/SP.J.1145.2013.00395]
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类球红细菌LHⅡβ-亚基/α-亚基融合蛋白吸收光谱分析()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
19卷
期数:
2013年03期
页码:
395-398
栏目:
研究论文
出版日期:
2013-06-25

文章信息/Info

Title:
Absorption Spectral Analysis of Fused β-subunit and α-subunit Protein of Light-harvesting Ⅱ Complexes of Rb. sphaeroides
作者:
丁国林胡宗利陈国平赵志平程丽静
(重庆大学生物工程学院 重庆 400044)
Author(s):
DING GuolinHU ZongliCHEN GuopingZHAO ZhipingCHENG Lijing
(College of Bioengineering, Chongqing University, Chongqing 400044, China)
关键词:
类球红细菌融合表达柔性序列捕光色素蛋白复合体Ⅱ近红外吸收光谱蛋白鉴定
Keywords:
Rb. sphaeroides fusion expression flexible sequence (FS) light-harvesting Ⅱ complex (LHⅡ) near infrared absorption spectral protein identification
分类号:
Q936
DOI:
10.3724/SP.J.1145.2013.00395
摘要:
类球红细菌(Rb. sphaeroides)捕光色素蛋白复合体Ⅱ(LHⅡ)由9对β-亚基和α-亚基组成,分别是pucB和pucA基因编码的. 为探究β-亚基和α-亚基融合后是否能够形成正常结构与功能的LHⅡ,在pucB和pucA基因之间加入一段柔性序列(Flexible sequence,FS),构建pucB-FS-pucA融合表达载体,导入双缺失突变体Rb. sphaeroides CQU68菌株中,经IPTG诱导,近红外700 -900 nm吸收光谱扫描细菌活体,亲和层析分离纯化蛋白并进行SDS-PAGE电泳鉴定. 与对照(未融合的菌株)相比,融合菌株在800 nm和850 nm附近也出现了两个典型的特征吸收峰,但吸收峰值略微降低并蓝移,电泳结果说明融合蛋白在类球红细菌中获得成功表达. 可见,两亚基融合后能够形成有活性的LHⅡ并进入到光合膜系统.
Abstract:
The light-harvesting Ⅱ complex (LHⅡ) of Rb. sphaeroides is composed of nine β-subunits and α-subunits, encoded by pucB gene and pucA gene. This study aimed to verify whether fused β-subunit and α-subunit still have the ability to form LHⅡ with normal structure and function. A flexible sequence (FS) was inserted between pucB gene and pucA gene, and the expression vector pucB-FS-pucA was constructed and transferred to the dual-deletion mutant Rb. sphaeroides CQU68 strain. It was then induced by IPTG, scanned by 700-900 nm near infrared absorption spectral, and finally identified by protein SDS-PAGE electrophoresis of affinity chromatography. Compared to the control (unfused strain), the fused strain also showed two typical absorption peaks at 800 nm and 850 nm, though slightly lower with blue shift. The electrophoresis showed a successful expression of the fusion protein in Rb. sphaeroides. In summary, the results suggested that the fusion protein can form an active LHⅡ and enter the photosynthetic membrane system.

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备注/Memo

备注/Memo:
国家自然科学基金项目(31100089)和中央高校基本科研业务费项目(CDJXS10232209)资助
更新日期/Last Update: 2013-06-20