gloeobacter violaceus pcc 7421藻胆蛋白裂合酶的蛋白质工程研究 word格式.docxVIP

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gloeobacter violaceus pcc 7421藻胆蛋白裂合酶的蛋白质工程研究 word格式.docx

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gloeobacter violaceus pcc 7421藻胆蛋白裂合酶的蛋白质工程研究 word格式

AbstractPhycobilisomes serve as the light-harvesting antenna in cyanobacteria and red algae. These supramolecular protein complexes, are primarily composed of phycobiliproteins, a brilliantly colored family of proteins bearing covalently attached, open-chain tetrapyrroles known as phycobilins. In additional, phycobilisomes also contain smaller amounts of linker peptides, which are required for proper assembly and functional organization of phycobilisomes. Phycobilin chromophores are generally bound to apoprotein at conserved positions by cysteinyl thioether linkages to generate phycobiliproteins. In vivo, the correct attachment of most chromophores is catalyzed by binding-site and chromophore-specific lyases.Lyases of PEB which binding to phycoerythrin subunits has not been all found. 20 genes in Gloeobacter violaceus PCC 7421 which have been analyzed by blast search have been reconctructed in order to seek the correct lyase by the method of in vivo reconstitution. This paper constructed gene glr1614 by applying the technique of molecular cloning and accomplished the construction of relevant plasmids, such aspACYCDuet-ho1-gll1188 、 pACYCDuet-ho1-glr1257 、 pACYCDuet-ho1-glr1259 、pCDFDuet-glr1192, etc. This work will facilitate further research on these lyases.In this paper, by using the method of reconstitution in vitro and SDSelectrophoresis, no obvious evidence is obtained indicating protein complex between CpeS2, MpeU, NblB3, and CpeZ2, respectively, is formed. This paper constructs pBT-cpeB which could be applied in the bacterial two-hybrid system and further the study on phycobiliprotein lyases interactions in PCC7421.Keywords:phycobiliproteinphycoerythrinlyaseprotein complex目录摘要 IAbstract II1前言1.1藻胆体 (1)1.2藻胆蛋白 (3)1.3藻胆色素 (3)1.4藻胆蛋白与藻胆色素连接机制的研究现状 (5)1.5研究意义及展望 (7)2基因 glr1614 的克隆与表达2.1引言 (9)2.2材料、试剂与仪器 (9)2.3实验方法 (11)2.4结果讨论 (17)3质粒构建 (20)3.1pACYCDuet-ho1-gll1188、pACYCDuet-ho1-glr1257、 pACYCDuet-ho1-glr1259 的质粒构建3.2pCDFDuet-glr1192 的质粒构建 (27)3.3pE