Studies On The Heterologous Expression Of LHI PufBA In Photosynthetic Bacteria Thermochromatium Tepidum

Photosynthetic bacteria is one of the earliest photosynthetic system of the general survival prokaryotes in nature.Thermochromatium(Tch.)Tepidum and Rhodospirillum(Rs.)Rubrum belong to the purple Photosynthetic bacteria and they are the gram-negative bacteria.The light-harvesting pigment protein complex of Thermochromatium(Tch.)Tepidum is composed of a reaction center and a Light Harvesting Complex.Because of the specificity of light-harvesting LHI structure,its the spectral scanning absorption peak appears at 915 nm,which is 880 nm with other Photosynthetic bacteria are very different.Rhodospirillum(Rs.)RubrumH2 is a mutant strain of the gene about light reaction center and light-harvesting pigment protein mutation.In this way,Rs.rubrumH2 can be successfully expressed and transported to the cell membrane by the structural specificity of ICM when the foreign gene is successfully inserted into the light reaction center and the light harvesting protein.In this paper,the gene pufBA,which encodes the light-harvesting protein LHI,was successfully cloned by in vitro gene amplification.The recombinant LHI was successfully cloned into the vector pET28 a,and the target protein LHI was expressed and purified successfully.The LHI polyclonal antibody was successfully obtained by immunizing the mice.The recombinant LHI gene which is the pufBA gene was cloned into the pPUCterm vector and transformed into the mutant strain Rs.rubrumH2 by the combination of WM3064 and E.coli.The light-harvesting protein LHI was successfully induced by hypoxia.The successful expression of the heterologous protein LHI was confirmed by the method of spectral scanning,and its expression position was confirmed on the cell membrane of Rs.rubrumH2.The successful expression of the heterologous membrane protein of light-harvesting pigment LHI which comes from Photosynthetic bacteria Thermochromatium(Tch.)Tepidum is helpful for revealing the specificity of the light-harvesting pigment LHI of Thermochromatium(Tch.)Tepidum,and its heterologous membrane protein expression Which can be used to reveal the photosynthesis of Photosynthetic bacteria,and to make better use of light energy,and to provide important theoretical basis for the research of membrane protein expression system Rs.rubrumH2.