| Combinational biosynthesis and characterization of a fluorescent 82 beta-phycocyanin of Spirulina platensis | |
Guan XiangYu2; Zhang WeiJie3; Chi XiaoYuan3; Lin HanZhi3,4; Wang JinFeng3; Qin Song1
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| 发表期刊 | CHINESE SCIENCE BULLETIN
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| ISSN | 1001-6538 |
| 2012-09-01 | |
| 卷号 | 57期号:25页码:3294-3299 |
| 关键词 | Escherichia Coli Phycocyanin Fluorescence Recombinant |
| 产权排序 | [Qin Song] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China; [Guan XiangYu] China Univ Geosci, Sch Ocean Sci, Beijing 100083, Peoples R China; [Zhang WeiJie; Chi XiaoYuan; Lin HanZhi; Wang JinFeng] Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China; [Lin HanZhi] Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China |
| 通讯作者 | Qin, S (reprint author), Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China.,sqin@yic.ac.cn |
| 作者部门 | 海岸带生物学与生物资源利用所重点实验室 |
| 英文摘要 | To biosynthesize fluorescent Spirulina platensis (Sp) beta-phycocyanin (PC) in Escherichia coli, a BLASTP search for homologs of the cpeS gene, a chromophore lyase, was performed against the Synechocystis sp. PCC 6803 (S6) proteome. A highly homologous gene, slr2049, was obtained from the S6 genome. Sites 82 and 153 in beta-phycocyanin of Sp were modified by site-directed mutagenesis. Two recombinant expression vectors were constructed and transformed into E. coli BL21: (i) pCDF-cpcB (C153A)-slr2049-sll0583-ho1-pcyA; and (ii) pCDF-cpcB (C82I)-slr2049-sll0583-ho1-pcyA. Lyases encoded by the genes slr2049 and sll0583 catalyzed the linking of Sp 82 beta-PC to phycocyanobilin (PCB), and fluorescent CpcB (C153A)-PCB was generated. We present a strategy for the co-expression of multiple genes in a single expression vector to identify the function of an unknown gene. Recombinant phycobiliproteins produced on a large scale are promising fluorescent tags for diagnostics and pharmacology.; To biosynthesize fluorescent Spirulina platensis (Sp) beta-phycocyanin (PC) in Escherichia coli, a BLASTP search for homologs of the cpeS gene, a chromophore lyase, was performed against the Synechocystis sp. PCC 6803 (S6) proteome. A highly homologous gene, slr2049, was obtained from the S6 genome. Sites 82 and 153 in beta-phycocyanin of Sp were modified by site-directed mutagenesis. Two recombinant expression vectors were constructed and transformed into E. coli BL21: (i) pCDF-cpcB (C153A)-slr2049-sll0583-ho1-pcyA; and (ii) pCDF-cpcB (C82I)-slr2049-sll0583-ho1-pcyA. Lyases encoded by the genes slr2049 and sll0583 catalyzed the linking of Sp 82 beta-PC to phycocyanobilin (PCB), and fluorescent CpcB (C153A)-PCB was generated. We present a strategy for the co-expression of multiple genes in a single expression vector to identify the function of an unknown gene. Recombinant phycobiliproteins produced on a large scale are promising fluorescent tags for diagnostics and pharmacology. |
| 文章类型 | Article |
| 资助机构 | Jiangsu Provincial Marine Biotechnology Key Lab Open Foundation[2010HS02]; National Natural Science Foundation of China[41006094] |
| 收录类别 | SCI |
| 语种 | 英语 |
| 关键词[WOS] | SYNECHOCOCCUS SP PCC-7002 ; BLUE-GREEN-ALGAE ; C-PHYCOCYANIN ; ESCHERICHIA-COLI ; ALPHA-PHYCOCYANIN ; FERREDOXIN OXIDOREDUCTASE ; CHROMOPHORE ATTACHMENT ; ANTIOXIDANT ACTIVITY ; EXPRESSION ; LYASE |
| 研究领域[WOS] | Science & Technology - Other Topics |
| WOS记录号 | WOS:000307750500007 |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.yic.ac.cn/handle/133337/6096 |
| 专题 | 海岸带生物学与生物资源利用重点实验室_海岸带生物学与生物资源保护实验室 |
| 作者单位 | 1.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China 2.China Univ Geosci, Sch Ocean Sci, Beijing 100083, Peoples R China 3.Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China 4.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Guan XiangYu,Zhang WeiJie,Chi XiaoYuan,et al. Combinational biosynthesis and characterization of a fluorescent 82 beta-phycocyanin of Spirulina platensis[J]. CHINESE SCIENCE BULLETIN,2012,57(25):3294-3299. |
| APA | Guan XiangYu,Zhang WeiJie,Chi XiaoYuan,Lin HanZhi,Wang JinFeng,&Qin Song.(2012).Combinational biosynthesis and characterization of a fluorescent 82 beta-phycocyanin of Spirulina platensis.CHINESE SCIENCE BULLETIN,57(25),3294-3299. |
| MLA | Guan XiangYu,et al."Combinational biosynthesis and characterization of a fluorescent 82 beta-phycocyanin of Spirulina platensis".CHINESE SCIENCE BULLETIN 57.25(2012):3294-3299. |
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