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Studies on Autoassemblying of Phycocyanin of Spirμlina Platensis and Functional Comparison of Some Phycocyanobilin Lyases

Author: YiJunJie
Tutor: ZhangXueCheng
School: Ocean University of China
Course: Genetics
Keywords: Spirulina platensis Phycocyanin Rice Autocatalytic Lyase Reorganization
CLC: Q946
Type: Master's thesis
Year: 2010
Downloads: 65
Quote: 0
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Abstract


The thesis Spirulina platensis (Spirμlina platensis FACHB314) genome as a template clone phycocyanin α subunit gene cpcA catalytic apoprotein color binding lyase gene cpcE and cpcF, together with the the ferrite protein redox the gene pcyA and the heme oxidase gene hoxl (template for the Synechocystis sp.PCC6803 genomic DNA), in E. coli the use of spirulina lyase catalytic synthesis of fluorescence activity phycocyanin α subunit . In the course of the study also found that phycocyanin α subunit can in no case lyase in combination with the phycocyanin choline autocatalytic, but the combination of inefficient, the fluorescence intensity of only phycocyanin lyase catalyzed of 34%. Clone the Spirulina platensis phycocyanin β subunit gene cpcB, Synechococcus Synechococcus sp PCC6803 lyase gene the PCC 7002 lyase gene cpcT of, cpcU and cpcS (7002), Synechocystis algae of Synechocystis sp. cpcS (6803). By building different expression strain compared lyase CpcT, the CpcU CpcS (7002), CpcU / CpcS (7002), CpcS (6803) phycocyanin p subunit catalysis effect. Experimental results show CpcU / CpcS (7002) as a heterodimer to effective catalytic phycocyanobilin proteins p subunit phycocyanobilin choline binding, and its expression strains fluorescence intensity of the supernatant was 110.1; CpcT catalytic efficiency minimum, the fluorescence intensity of only 52.71; the catalytic effect of the CPCS (6803) residing between the fluorescence intensity of 81.69. The phycocyanin p subunit can be combined with with the phycocyanin choline independently, and since the catalytic efficiency is relatively high, the fluorescence intensity of expression strains BUS (7002) close to 111.5. Building the expression strain AUS (7002), found that catalytic phycocyanin α subunit phycocyanin choline PCB Synechococcus lyase CpcU / CpcS (7002) can also be combined with fluorescence intensity lyase catalytic phycocyanin α subunit of 1.8 times. Clone Spirulina platensis phycocyanin groups are due to cpcBA, ussBcpcBA, α and β subunits are present in the case as well as in the case with phycocyanin promoter by various expression strain comparison lyase CpcE / CpcF, CpcU / CpcS (7002) the phycocyanin catalytic effect. The experimental results indicate that simultaneous expression of the αβ subunit expression strain protein synthesis effect is better than the effect of the α-subunit or β subunit expression strain containing only. Which express the best expression of the effect of the strain uBA-EF, the maximum emission peak of 640nm, the closest natural phycocyanin maximum emission peak (645nm) and fluorescence intensity (369.2). Suggesting can refer to the expression strain uBA-EF to build rice phycocyanin fluorescence activity expression system. This paper studies the self-catalysis phycocyanin and phycocyanin lyase function of comparative studies, the use of E. coli expression system to explore the phycocyanin fluorescence activity synthesis mechanism for expression in rice with fluorescent activity the phycocyanin provide theoretical and experimental basis.

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